假俭草体细胞抗寒突变体的获得及其SRAP分子鉴定

假俭草抗寒性差是限制其广泛应用的主要因素之一。本研究目标是以优良假俭草选系E-126为材料,经低温诱导和筛选获得体细胞抗寒突变体。结果表明,假俭草种子诱导的愈伤组织在继代培养的过程中,经0℃的低温条件下培养26 d,获得了2块存活的愈伤组织,该愈伤组织经过继代增殖后,进行分化、生根和移栽,获得株系1和株系2。苗期外部形态观察结果表明,假俭草低温诱导的株系1、株系2和对照在叶色、叶毛、叶长和叶宽上均没有显著性差异。半致死温度分析结果表明,株系1、株系2以及对照叶片半致死温度(LT₅₀)分别为-6.646,-6.546和-5.351℃,处理与对照之间差异显著,且都低于对照,但株系1和株系2之间无显著性差异。SRAP的结果表明,假俭草低温诱导的株系1和株系2在110,230以及240 bp处均存在相同特征带,表明假俭草体细胞突变体植株的变异稳定且在分子水平与对照存在差异,但株系1和株系2无差异。因此,株系1和株系2可作为同一体细胞抗寒突变体株系加以利用。     

High-throughput screening reveals small molecule modulators inhibitory to Acidovorax citrulli

Acidovorax citrulli is a seed-transmitted gram-negative bacterium that can cause substantial economic yield loss in watermelon and melon production worldwide. Four small-molecule libraries containing 4,952 compounds were selected for high-throughput screening against A. citrulli wild-type strain Xu3-14 by evaluation of growth inhibition. One hundred and twenty-seven molecules (2.5% hit rate) were identified as bactericidal or bacteriostatic against A. citrulli at 100 μM. Secondary screens indicated that 27 candidate compounds were more effective against A. citrulli Group II strains than Group I strains (classified using repetitive element PCR). Several compounds were inhibitory to other pathogenic bacteria, including Clavibacter michiganensis subsp. michiganensis and Xanthomonas campestris pv. campestris, but did not affect the growth of plant beneficial bacteria Pseudomonas fluorescens and Bacillus subtilis. More than half of the compounds did not inhibit germination of Arabidopsis or watermelon seeds. The effect of small molecules on A. citrulli seed-to-seedling transmission was evaluated by applying each compound to inoculated watermelon seeds and assessing seedling infection. Nine compounds were chosen for further investigation based on their reduction of percentage seedling infection and compiling scores on their specificity, sensitivity, and phytotoxicity obtained in the secondary screens. The five best compounds were selected (thiamphenicol, nadifloxacin, pipemidic acid, ciclopirox, and zinc pyrithione) for greenhouse tests and were found to effectively reduce the seed-to-seedling transmission of A. citrulli in both artificially and naturally infested seeds. These top five compounds provide a basis for future development of an A. citrulli-specific bactericide.     

草莓褐色轮斑病病原鉴定及室内药效研究*

草莓褐色轮斑病近年越来越严重,尤其在草莓育苗阶段。从草莓发病叶片、匍匐茎上分离得到病原菌,对病原菌进行形态特征观察、生物学特性研究、ITS序列分析以及室内药效试验。结果表明：该病原菌为Sphaeronaemella fragariae。该菌菌丝生长最适温度范围是25～28 ℃;适宜pH为6;在供试的几种碳、氮源中,最适的碳源是蔗糖,最适的氮源是酵母浸出液。在供试的9种药剂中,以咪鲜胺1 000倍液对病菌的抑制作用最好。     

Evaluation of an Indian collection of black gram germplasm and identification of resistance sources to Mungbean yellow mosaic virus

Yellow mosaic disease (YMD) of black gram is an important production constraint in India. Black gram germplasm, consisting of 344 accessions, originally collected from different geographic regions of India and conserved in National Gene Bank at National Bureau of Plant Genetic Resources, India, were evaluated for their response against YMD. None of the accessions tested was found to be immune, however, considerable variation in disease response was observed among them. Of the 344 accessions, 32 showed resistance in the field during the rainy season (June–September) 2010 as assessed by coefficient of infection (CI) and area under disease progress curve (AUDPC). These accessions were re-evaluated during rainy season (June–September) 2011 and 2012 for their disease resistance along with yield-associated characters. Eight accessions that exhibited consistently highly resistant (HR) or resistant (R) reaction (CI ≤ 9) along with lower AUDPC value (≤650), were further evaluated following inoculation with whiteflies or via agroinoculation. Along with the resistant trait, these eight accessions were also found either similar or even better in their yield potential as compared to check. After artificial inoculation using viruliferous whiteflies under glasshouse, of the eight field-resistant accessions, response of two (IC144901 and IC001572) were categorized as HR as <10% plants exhibited minute yellow specks after 17–22 dpi. Two accessions viz. IC011613 and IC485638 showed <20% incidence with bright yellow specks or spots after 15–20 dpi and were grouped into R category. The resistance of these four accessions was further confirmed through agroinoculation of the infectious cloned viral DNA under controlled conditions. In resistant accessions low accumulation of viral DNA was observed over a period of time. These four accessions could be used in future breeding programmes to develop black gram cultivars resistant to YMD or could be used directly as varieties to manage YMD after adaption to various agro-climatic regions.     

中兽药口服液中隐性药物筛查高分辨质谱方法的建立

建立了测定中兽药口服液中170种隐性药物的超高效液相色谱—四极杆静电场轨道阱质谱( HPLC- Q-Exactive Focus/MS) 高通量筛查方法。通过构建药物筛查数据库并与样品中未知物信息进行比对,从而实现快速筛查中兽药口服液中隐性药物的种类和含量。数据库包括磺胺类、喹诺酮类、β-受体激动剂类、硝基咪唑类、苯并咪唑类、蛋白同化激素类、糖皮质激素类、喹噁啉类、四环素类、抗病毒类、酰胺醇类和真菌毒素类等12类170种药物的筛查参数。样品中人为添加化学药物,经1%甲酸乙腈提取,QuEChERs净化,氮气吹干,0.2%甲酸水复溶上机,正负模式扫描,TraceFinder软件筛查定性,当药物添加浓度为400ng/g时,170种添加药物的检出率约为92.8%,随着药物浓度逐步提高至1 μg/g时,所有添加药物均可检出。结果表明,该方法可满足中兽药口服液中隐性药物高通量快速筛查的需要,为打击畜禽养殖投入品中非法使用化学药物的行为提高技术支持和数据支撑。     

A greenhouse screening technique to assess rust resistance in sorghum∗

Abstract

Studies were conducted to determine the influence of plant growth stage, inoculum density, temperature, and relative humidity (RH) on development of rust (Puccinia pupurea) in sorghum (Sorghum bicolor). Rust development was maximum (>80% severity), when plants of a susceptible sorghum genotype (IS 18420) were inoculated at the four‐ to five‐leaf stage with an inoculum concentration of 4 × 10⁶ urediniospores per ml and incubated at 20–25°C under high RH (>90%) for 24 h. Disease severity (percentage leaf area covered with rust pustules) scores were taken 2 weeks after inoculation. Using this technique, 29 sorghum genotypes were screened for rust resistance in a greenhouse. This technique proved effective In discerning resistant and susceptible genotypes, and IS 3979, ICSH 110, ICSH 86647 and ICSH 871035 were identified resistant (<20% rust severity) compared with a susceptible control IS 18420 (90% rust severity). This technique is simple and rapid, and can be used effectively and economically to screen, on a large scale, germplasm lines and breeding populations in the greenhouse.     

农业执法案件中未知粉末的分析研究

对农业执法案件中缴获的未知粉末进行分析研究。应用超高效液相色谱-飞行时间高分辨质谱（UPLC-TOF-HRMS）获得未知物分子离子峰的精确质量数、天然同位素信息,二级特征碎片的精确质荷比,并应用SCIEX PeakView及MasterView软件进行分析。经过靶向及非靶向筛查,同时参考文献报道,推导出未知粉末中的疑似化合物。采用高效液相色谱-二极管阵列检测器（HPLC-PDA）法及UPLC-TOF-HRMS法进行了双重确证。结果显示该未知粉末中含克仑特罗（16 g?kg-1）及其合成前体克仑特罗杂质B（455 g?kg-1）。本文详述的分析技术可为未知添加物的筛查和确证提供思路和技术参考。     

发酵饲料中抑菌菌种筛选及抑菌性能评估方法的初步研究

试验通过牛津杯法筛选出抑制金黄色葡萄球菌和大肠杆菌性能较强的一株乳酸菌L3和两株芽孢杆菌D9、K9.将其用于制作发酵饲料,提升发酵饲料的抑菌性能.试验初步研究发酵饲料的体外抑菌性能评估方法.结果表明,用80％的乙醇溶液作为发酵饲料的前提取溶剂可增加抑菌物质的溶解,浓缩提取液的浓度可使发酵样品的抑菌效果更加明显.     

Identification of novel juvenile-hormone signaling activators via high-throughput screening with a chemical library

Juvenile hormone (JH) is an insect-specific hormone that regulates molting and metamorphosis. Hence, JH signaling inhibitors (JHSIs) and activators (JHSAs) can be used as effective insect growth regulators (IGRs) for pest management. In our previous study, we established a high-throughput screening (HTS) system for exploration of novel JHSIs and JHSAs using a Bombyx mori cell line (BmN_JF&AR cells) and succeeded in identifying novel JHSIs from a chemical library. Here, we searched for novel JHSAs using this system. The four-step HTS yielded 10 compounds as candidate JHSAs; some of these compounds showed novel basic structures, whereas the others were composed of a 4-phenoxyphenoxymethyl skeleton, the basic structure of several existing JH analogs (pyriproxyfen and fenoxycarb). Topical application of seven compounds to B. mori larvae significantly prolonged the larval period, suggesting that the identified JHSAs may be promising IGRs targeting the JH signaling pathway.     

抗弓形虫和新孢子虫药物体外筛选方法的研究进展

弓形虫和新孢子虫是2种在形态结构及生物学特征方面非常相似的细胞内寄生性原虫,二者引起的疾病对人和动物健康造成了严重危害。开发和筛选高效、低毒的抗弓形虫和新孢子虫药物一直是抗原虫药物研究领域的重要方向之一。药物筛选包括体外筛选和体内筛选,体外筛选的结果是进行体内筛选的依据和基础。对抗弓形虫和新孢子虫药物的体外筛选方法研究进展进行综述,以期为开发更加高效的抗弓形虫和新孢子虫药物体外筛选技术提供参考。