ACE inhibition in goats under fixed‐time artificial insemination protocol increases the pregnancy rate and twin births

To assess the effect of the angiotensin‐converting enzyme (ACE) inhibition on the efficiency of the fixed‐time artificial insemination (TAI), 69 goats were divided randomly into two groups: enalapril (n = 35) and control (n = 34). In the experiment, all animals underwent the protocol of fixed‐time artificial insemination for 12 days. Enalapril group received enalapril maleate dissolved in saline (Enalapril, Lab Teuto Ltda) subcutaneously at the following doses: 0.2 mg/kg/day in D0‐D2; 0.3 mg/kg/day in D3‐D6 and 0.4 mg/kg/day in D7‐D11. The control group received the corresponding volume of 0.9% saline solution. We performed a single insemination 36 hr after sponge removal using frozen semen from two adult male goats with recognized fertility. The ultrasound pregnancy diagnosis was 30 days after the artificial insemination (AI). There was significant increase in pregnancy rates and twinning as well as a decrease in foetal loss in animals receiving enalapril (p < .01). The use of ACE inhibitors during the TAI protocol was shown to be a promising alternative to increase the efficiency of such reproductive biotechnology.     

Comparative bioactivity of selected seed extracts from Brazilian Annona species and an acetogenin-based commercial bioinsecticide against Trichoplusia ni and Myzus persicae

The acute and chronic toxicity of ethanolic seed extracts from selected Brazilian Annona species (Annona montana Macfadyen, Annona mucosa Jacquin, Annona muricata Linnaeus, and Annona sylvatica A. St.-Hil) and an acetogenin-based commercial bioinsecticide (Anosom^®) were investigated against the cabbage looper Trichoplusia ni Hübner (Lepidoptera: Noctuidae) and the green peach aphid Myzus persicae (Sulzer) (Hemiptera: Aphididae). In the laboratory, extracts of A. mucosa and A. sylvatica as well as Anosom^® were especially active through oral and topical administration. A greenhouse trial showed that a formulated A. mucosa extract and Anosom^® were highly effective (>98% mortality) against third instar T. ni larvae, and comparable to a pyrethrin-based commercial insecticide (Insect Spray^®) used as a positive control. Similar to results with T. ni, A. mucosa extract showed the greatest aphicidal activity followed by A. sylvatica extract and Anosom^®. In another greenhouse trial, aphid population reduction from the formulated A. mucosa extract was superior to that provided by other treatments including the positive control. Though inferior to the A. mucosa extract, the acetogenin-based commercial insecticide (Anosom^®) and A. sylvatica extract also reduced aphid populations in a manner comparable to the positive control. Botanical insecticides based on these Annonaceae derivatives could be useful in the framework of Brassica IPM in Brazil and elsewhere, especially for organic production.     

Effects of natural protease inhibitors on high protease activity flours

Wheat grain damaged by wheat bug (Eurygaster spp.) contains the bug salivary secretion which hydrolyses the gluten needed for the dough quality of breadmaking due to its proteolytic activity. Since the protease inhibitors are widespread throughout the plant kingdom, the possibility of using some plants, especially food and feed legumes to decrease the proteolytic activity of flours milled from bug damaged wheats was investigated. The proteolytic activity was considerably inhibited by pefabloc-SC and EDTA-Na₂, suggesting that bug protease(s) included serine and metallo proteases. Extracts from cones of hops, seeds of grass pea, red kidney bean and sunflower caused reduction in the activity of bug protease(s). Effects of hop extract on electrophoretic, rheological properties of high protease activity flours milled from different bread wheat cultivars damaged by the bug were also studied. The dough development time and stability values of high protease activity flours increased considerably with hop extract at the lowest addition level (10:1, flour to hop extract ratio). The doughs supplemented with the hop extract had higher maximum resistance and lower extensibility values as compared to their controls. These results suggested that hop extract had improving effects on high protease activity flours due to the bug damage.     

黄瓜低霜霉威残留性相关基因SDH克隆及表达

通过Solexa技术和比较基因组学方法鉴定霜霉威胁迫下黄瓜果实差异表达基因,筛选黄瓜低霜霉威残留性相关基因SDH。研究利用PCR克隆获得SDH基因全长,命名为Cs SDH。构建亚细胞定位表达载体,利用农杆菌浸润法注射烟草叶片后激光共聚焦显微镜下观察;分别用荧光定量RT-PCR方法和酶联免疫法研究高、低农残品系D9320和D0351中Cs SDH基因在农药霜霉威处理后,不同时间点、不同组织部位时空表达特征和琥珀酸脱氢酶活性。结果表明,Cs SDH基因在烟草叶片细胞中被定位在细胞膜上。黄瓜低农残品种D0351中,Cs SDH基因在果实受霜霉威胁迫后反应迅速,主要在处理前期响应表达强烈,琥珀酸脱氢酶活性显著增加,且该基因表达量显著高于其在高农残品种D9320中表达量。Cs SDH果、叶中表达量较茎中高,且各组织部位表达量高低顺序稳定,为叶果茎。黄瓜高农残品种D9320中,Cs SDH基因表达量在0.5、1、3、12 h呈现上调表达,6、9 h基因表达量差异不显著,48 h出现上调表达高峰。说明克隆得到的Cs SDH基因属农药处理前期响应基因,且该基因可能在降低黄瓜农药残留过程中发挥重要作用。试验通过研究黄瓜琥珀酸脱氢酶基因(SDH)在霜霉威胁迫下表达情况,为挖掘黄瓜低农药残留性功能基因以及探明其作用分子机制奠定基础。     

Molecular cloning,purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein

The Brucella mdh gene was successfully cloned and expressed in E. coli. The purified recombinant malate dehydrogenase protein (rMDH) was reactive to Brucella-positive bovine serum in the early stage, but not reactive in the middle or late stage, and was reactive to Brucella-positive mouse serum in the late stage, but not in the early or middle stage of infection. In addition, rMDH did not react with Brucella-negative bovine or mouse sera. These results suggest that rMDH has the potential for use as a specific antigen in serological diagnosis for early detection of bovine brucellosis.     

Genetic basis and origin of resistance to acetolactate synthase inhibitors in Amaranthus palmeri from Spain and Italy

BACKGROUND

Amaranthus palmeri is an aggressive annual weed native to the United States, which has become invasive in some European countries. Populations resistant to acetolactate synthase (ALS) inhibitors have been recorded in Spain and Italy, but the evolutionary origin of the resistance traits remains unknown. Bioassays were conducted to identify cross-resistance to ALS inhibitors and a haplotype-based genetic approach was used to elucidate the origin and distribution of resistance in both countries.

RESULTS

Amaranthus palmeri populations were resistant to thifensulfuron-methyl and imazamox, and the 574-Leu mutant ALS allele was found to be the main cause of resistance among them. In two Spanish populations, 376-Glu and 197-Thr mutant ALS alleles were also found. The haplotype analyses revealed the presence of two and four distinct 574-Leu mutant haplotypes in the Italian and Spanish populations, respectively. None was common to both countries, but some mutant haplotypes were shared between geographically close populations or between populations more than 100 km apart. Wide genetic diversity was found in two very close Spanish populations.

CONCLUSION

ALS-resistant A. palmeri populations were introduced to Italy and Spain from outside Europe. Populations from both countries have different evolutionary histories and originate from independent introduction events. ALS resistance then spread over short and long distances by seed dispersal. The higher number and genetic diversity among mutant haplotypes from the Spanish populations indicated recurrent invasions. The implementation of control tactics to limit seed dispersal and the establishment of A. palmeri is recommended in both countries. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

植物表达载体pGMAR-BADH的构建

将甜菜碱醛脱氢酶基因BADH和植物表达载体pGMAR通过内切酶BamHI和KpnI双酶切,T4连接酶进行连接反应。重组质粒在大肠杆菌菌株DH5α内扩增,提取并纯化质粒。经鉴定,基因BADH已被完整、正确的插入到pGMAR载体中,并成功将BADH插入到载体pGMAR的两个MAR序列之间。     

Quinone Binding Sites of Membrane Proteins as Targets for Inhibitors

Quinones are a central component of most photosynthetic and respiratory electron transfer chains. The proteins that reduce and oxidise these quinones have quinone binding sites—Q sites—that are good targets for pesticides. This paper reviews the diversity of these sites, their possible structure, and the types of compounds that act upon them.