Geographical cline of DNA variation within the F intersterility group of Heterobasidion annosum in Italy

Eighty-six Heterobasidion annosum isolates, mainly belonging to the F intersterility group and obtained from 32 different geographical localities in Italy, were subjected to genetic analysis by the Random Amplified Polymorphic DNA (RAPD) markers. The similarity between F and S groups was higher than that between F and P. In UPGMA Cluster Analysis, the F isolates originating from the same locality usually grouped in the same cluster. The isolates also showed a tendency to group at the level of larger geographical areas. Within the F group, isolates from the south of the Italian peninsula showed the highest genetic variation and northern isolates from the Alpine regions showed the lowest. This indicates a gradual cline along the peninsula. The genetic variability in the Italian F group is discussed in relation to the past and present distribution of the host species in Italy and Europe.     

Chemical comparison of goldenseal (Hydrastis canadensis L.) root powder from three commercial suppliers

The characterization of herbal materials is a significant challenge to analytical chemists. Goldenseal (Hydrastis canadensis L.), which has been chosen for toxicity evaluation by NIEHS, is among the top 15 herbal supplements currently on the market and contains a complex mixture of indigenous components ranging from carbohydrates and amino acids to isoquinoline alkaloids. One key component of herbal supplement production is botanical authentication, which is also recommended prior to initiation of efficacy or toxicological studies. To evaluate material available to consumers, goldenseal root powder was obtained from three commercial suppliers and a strategy was developed for characterization and comparison that included Soxhlet extraction, HPLC, GC-MS, and LC-MS analyses. HPLC was used to determine the weight percentages of the goldenseal alkaloids berberine, hydrastine, and canadine in the various extract residues. Palmatine, an isoquinoline alkaloid native to Coptis spp. and other common goldenseal adulterants, was also quantitated using HPLC. GC-MS was used to identify non-alkaloid constituents in goldenseal root powder, whereas LC-MS was used to identify alkaloid components. After review of the characterization data, it was determined that alkaloid content was the best biomarker for goldenseal. A 20-min ambient extraction method for the determination of alkaloid content was also developed and used to analyze the commercial material. All three lots of purchased material contained goldenseal alkaloids hydrastinine, berberastine, tetrahydroberberastine, canadaline, berberine, hydrastine, and canadine. Material from a single supplier also contained palmatine, coptisine, and jatrorrhizine, thus indicating that the material was not pure goldenseal. Comparative data for three commercial sources of goldenseal root powder are presented.     

甜菜SSR反应体系优化及重要农艺性状分子标记

[目的]为选育优质甜菜新品种,利用SSR法筛选与高糖、高产、耐盐相关的分子标注.[方法]研究对甜菜SSR-PCR反应体系进行优化,并利用SSR分子标记方法和分离群体分组分析法(Bulked SegregateAnalysis,BSA)对具有高糖/低糖、低产/高产、耐盐/不耐盐三种重要农艺性状的甜菜亲本和F2代植株进行分析.[结果]研究建立了适宜甜菜的SSR-PCR反应体系为:20μL反应体系中含l×Buffer、2.0 mmol/LMg2+、1.5 uTaq DNA聚合酶、0.20 mmol/L dNTP、1.5 μmol/L引物、60 ng DNA模板.依据优化体系,对不同农艺性状的甜菜亲本与F2代进行SSR-PCR扩增分析,高糖性状获得了200和100 bp两条与高糖性状连锁的标记,250和230 bp两条与高产性状连锁的标记,550、250和100 bp三条与耐盐性状紧密连锁的分子标记.[结论]研究获得的7条特异条带是与甜菜重要农艺性状连锁的分子标记,将为甜菜的育种工作提供重要的理论基础.     

绵羊LDHβ基因的生物信息学分析及其在不同剩余采食量绵羊肝脏和肌肉组织中的表达

【目的】为鉴定绵羊乳酸脱氢酶β(lactate dehydrogenase B,LDHβ)基因的分子特征,研究其在不同剩余采食量绵羊肝脏和肌肉组织中的表达差异.【方法】测定了137只‘湖羊’公羔的剩余采食量(residual feed intake,RFI),按RFI进行排序,分别筛选出RFI最高(high-residual feed intake,H-RFI)和RFI最低(low-residual feed intake,L-RFI)的羊各15只,屠宰后,采集肝脏和肌肉组织,利用Q-PCR技术检测绵羊LDHβ基因分别在H-RFI和L-RFI羊肝脏和肌肉中表达量,并利用生物信息学软件构建了该基因的系统进化树和预测其结构与功能.【结果】绵羊LDHβ基因开放阅读框(open reading frame,ORF)为1 005bp,编码334个氨基酸,其蛋白质分子质量为36 479.43U,理论等电点为6.40.绵羊LDHβ基因与山羊的亲缘关系较近,其次为牛,序列同源性高.功能结构域预测结果显示,该基因编码产物在内质网中参与辅酶因子生物合成的可能性最高.Q-PCR结果显示,绵羊LDHβ-mRNA基因在L-RFI羊肝脏和肌肉中表达量均显著低于H-RFI羊(P0.01).【结论】绵羊LDHβ基因作为能量代谢的关键酶参与绵羊饲料效率的调控.     

To pool or not to pool? Guidelines for pooling samples for use in surveillance testing of infectious diseases in aquatic animals

Samples from multiple animals may be pooled and tested to reduce costs of surveillance for infectious agents in aquatic animal populations. The primary advantage of pooling is increased population‐level coverage when prevalence is low (<10%) and the number of tests is fixed, because of increased likelihood of including target analyte from at least one infected animal in a tested pool. Important questions and a priori design considerations need to be addressed. Unfortunately, pooling recommendations in disease‐specific chapters of the 2018 OIE Aquatic Manual are incomplete and, except for amphibian chytrid fungus, are not supported by peer‐reviewed research. A systematic review identified only 12 peer‐reviewed aquatic diagnostic accuracy and surveillance studies using pooled samples. No clear patterns for pooling methods and characteristics were evident across reviewed studies, although most authors agreed there is a negative effect on detection. Therefore, our purpose was to review pooling procedures used in published aquatic infectious disease research, present evidence‐based guidelines, and provide simulated data examples for white spot syndrome virus in shrimp. A decision tree of pooling guidelines was developed for use by peer‐reviewed journals and research institutions for the design, statistical analysis and reporting of comparative accuracy studies of individual and pooled tests for surveillance purposes.     

不同处理方法对美国截叶胡枝子种子发芽的影响

美国截叶胡枝子的硬实率很高,发芽率只有17.3%.经过种子预处理,如采用不同温度下浸种、98%浓硫酸浸泡、机械破皮和机械破皮结合热水浸种等处理,结果表明,机械破皮后用60℃热水浸种处理的效果最好,发芽势和发芽率分别达到64.0%和74.3%;其次是机械破皮,平均发芽率达到50%;浓硫酸浸种可以显著提高种子发芽势,浓硫酸浸种5min效果最好,发芽率达到44.7%;热水浸种的处理效果不佳,最好的90℃热水浸种处理发芽率也仅有26.7%.