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.     

鸡粪中雌激素的检测及微生物降解

为检测鸡饲料和鸡粪中雌激素，优化其微生物降解条件，应用高效液相色谱（HPLC）法检测蛋鸡饲料和鸡粪中的雌激素，利用11种高效降解雌激素的单一菌株和等体积混合得到的菌群开展雌激素的降解实验。结果表明：只有蛋鸡鸡粪中含有雌二醇（E2），含量为230 ng·g^-1。微生物菌群的降解率高于单一菌株，在27℃下，10 d内能够降解76%的E2。研究表明，蛋鸡鸡粪中有雌激素存在，微生物菌群对雌激素的降解率高于单一菌株。     

A computer vision system for oocyte counting using images captured by smartphone

This work proposes a computer vision procedure for counting Twospot astyanax (Astyanax bimaculatus) oocytes in Petri dishes using images captured by smartphone. First, the proposed procedure uses simple linear iterative clustering (SLIC) to divide the images into groups of pixels (superpixels). Then, based on their color and space characteristics, the images are classified into light background, dark background, dirt, or oocyte by a machine learning algorithm. Five different types of machine learning algorithms were tested: support vector machines (SVM), decision trees using the algorithm J48 and random forest, k-nearest neighbors (k-NN), and Naive Bayes. To train the algorithms, 8.578 superpixels were classified by an expert into oocyte (n = 354), dirtiness (n = 651), dark background (n = 3.622), and light background (n = 3.951). Of the five learning algorithms, SVM obtained the best result with 97% correct oocyte recognition. Given the wide availability of smartphones, we therefore conclude that the presented procedure can be a valuable tool in future experiments and studies on fertilization and hatching success in Twospot astyanax.     

磁处理改善豌豆根际土壤生物学性状

采用盆栽的方法,分析磁处理对豌豆根际土壤生物学性状的影响,旨在探究磁处理对豌豆根际土壤肥力及健康状况的影响。试验设置无处理（对照CK）,添加铁粉（A）、磁化铁粉（B）、场强为0.2特斯拉（T）磁铁+铁粉（C）以及0.2 T磁铁（D）的处理,基于高通量测序技术分析磁处理对豌豆根际土壤生物学性状的影响。结果表明：C处理显著增加豌豆根际土壤中可培养细菌和真菌数量,同时微生物生物量C、N、P与CK相比分别增加了34.0%、46.0%、44.2%;β-葡萄糖苷酶、氨肽酶、磷酸酶酶活性分别增加了24.6%、23.7%、31.8%;另一方面,C处理中土壤细菌丰富度和多样性指数Ace、Chao1、Shannon指数显著高于CK;属（Genus）分类水平韦恩图分析中上,CK处理特有的属细菌主要有3个：Sneathiella、Desulfurispora、nannocystaceae;A处理特有的属细菌有7个;B处理特有的属细菌亦有7个;C处理特有的属细菌有11个;D处理特有的属细菌有8个。综上所述,C处理能够显著提高豌豆根际土壤中可培养微生物数量、微生物生物量（C、N、P）、酶活性和细菌多样性等多项指示土壤肥力和健康状况的生物学指标,表明磁场强度为0.2 T的磁铁+5 g磁化铁粉处理具有提高土壤肥力抵御土传病害的能力。     

茉莉酸甲酯诱导辣椒抗青枯病与活性氧代谢的关系

为探究茉莉酸甲酯(Methyl Jasmonate,Me JA)诱导辣椒抗青枯病效应与活性氧代谢相关酶的关系,以辣椒易感青枯病品种‘粤红1号’和抗青枯病品种‘辛香8号’幼苗为试验材料,在0.1mmol·L-1 Me JA喷雾处理后12、24、48、72和96 h接种青枯劳尔氏菌(Ralstonia solanacearum),对其进行青枯病病情指数与活性氧代谢相关酶——超氧化酶歧化酶(Superoxide Dismutase,SOD)、过氧化氢酶(Catalase,CAT)、过氧化物酶(Peroxidase,POD)和抗坏血酸过氧化物酶(Ascorbate Peroxidase,APX)活性以及丙二醛(Malondialdehyde,MDA)含量测定及相互关系的分析。结果表明:‘粤红1号’和‘辛香8号’的病情指数随喷Me JA处理时间的推移表现为先降后升,但都低于对照;而诱导效果则相反,Me JA处理对两个辣椒品种幼苗抗性的最适诱导时间均为接种前48 h。接种后0~96 h,Me JA喷雾处理的辣椒幼苗叶片SOD、CAT、POD和APX酶活性均显示先升后降的趋势,且明显高于对照,接种后24~48 h达到最高,而MDA含量则明显低于对照。因此,Me JA可诱导辣椒幼苗抗青枯病,其实现途径可能与提高活性氧代谢相关酶活性和缓解膜脂过氧化有关。     

Strong functional stability of soil microbial communities under semiarid Mediterranean conditions and subjected to long-term shifts in baseline precipitation

We investigated the effect of soil microclimate on the structure and functioning of soil microbial communities in a Mediterranean Holm-oak forest subjected to 10 years of partial rain exclusion manipulations, simulating average drought conditions expected in Mediterranean areas for the following decades. We applied a high throughput DNA pyrosequencing technique coupled to parallel measurements of microbial respiration (R_H) and temperature sensitivity of microbial respiration (Q₁₀). Some consistent changes in the structure of bacterial communities suggest a slow process of community shifts parallel to the trend towards oligotrophy in response to long-term droughts. However, the structure of bacterial communities was mainly determined by short-term environmental fluctuations associated with sampling date (winter, spring and summer) rather than long-term (10 years) shifts in baseline precipitation. Moreover, long-term drought did not exert any chronic effect on the functioning of soil microbial communities (R_H and Q₁₀), emphasizing the functional stability of these communities to this long-term but mild shifts in water availability. We hypothesize that the particular conditions of the Mediterranean climate with strong seasonal shifts in both temperature and soil water availability but also characterized by very extreme environmental conditions during summer, was acting as a strong force in community assembling, selecting phenotypes adapted to the semiarid conditions characterizing Mediterranean ecosystems. Relations of climate with the phylogenetic structure and overall diversity of the communities as well as the distribution of the individual responses of different lineages (genera) to climate confirmed our hypotheses, evidencing communities dominated by thermotolerant and drought-tolerant phenotypes.     

猪胆囊中沙门菌L型的分离培养与invA基因检测

为了解猪胆囊中沙门菌L型携带情况,在贵阳市屠宰场采集970例健康生猪的胆囊组织与胆汁标本,用常规细菌学方法和非高渗分离培养法分离沙门菌及其细菌L型,用PCR和核酸序列分析方法对稳定L型纯培养物进行沙门菌的invA基因检测。结果显示,970例生猪胆囊标本未检出沙门菌细菌型,细菌L型检出率为8.25%;80例细菌L型分离物中有50例invA检测阳性,占5.15%;占细菌L型阳性分离物62.50%。研究结果为生猪胆囊沙门菌L型感染的流行病学及其检查提供了依据。     

白叶枯病菌效应子XopN在拥有OsSWEET11同源基因的水稻品种上发挥毒性作用

【目的】黄单胞菌细胞外泌蛋白质(Xanthomonas outer proteins, Xops)是植物病原黄单胞菌高度保守的毒性效应子或毒性辅助组分,通过细菌Ⅲ型分泌系统分泌到细菌细胞外部,然后转入植物细胞而发挥病理作用。水稻黄单胞菌水稻致病变种即水稻白叶枯病菌的标准菌株PXO99^A分泌的XopN是一种毒性效应子,通过影响寄主免疫反应而使水稻发病。但是,XopN对病菌毒性的影响是否因水稻品种的不同而异,还有待研究。【方法】利用同源双交换技术,先敲除了PXO99^A的XopN基因,获得了∆XopN突变体,又经过遗传互补,得到了回补菌株∆XopN/XopN。通过营养肉汤液体培养,测定了XopN对病菌繁殖能力的影响;根据文献选用14个水稻品种,通过接种实验,测定了PXO99^A对这些品种的毒性与XopN敲除或回补的影响;在XopN发挥毒性作用的水稻品种上,测定了隐性抗病基因OsSWEET11/xa13与显性感病基因OsSWEET11/Xa13受病菌侵染而表达的情况,分析了XopN敲除或回补的影响。【结果】在营养肉汤液体培养过程中,病菌突变体菌株∆XopN的繁殖速度明显低于野生型PXO99^A。PXO99^A、∆XopN和∆XopN/XopN接种水稻后,根据其在水稻叶片组织内的繁殖量及随后产生的白叶枯病症状的严重程度,将供试的14个水稻品种分为两种情况。一是感病程度与病菌XopN是否敲除或回补无关,这有10个水稻品种（IRBB1、IRBB3、IRBB8、IRBB10、IRBB14、IR24、IRBB203、IRBB204、IRBB205和IRBB211）,PXO99^A、∆XopN和∆XopN/XopN对它们的毒性无明显差别。二是XopN对病菌毒性发挥作用的水稻品种,包括高度感病的3个品种（IRBB208、Asominori和日本晴）和低感品种IRBB13。与PXO99^A或∆XopN/XopN相比,∆XopN对这4个水稻品种的毒性大为降低。在IRBB13上,病菌侵染对隐性抗病基因OsSWEET11/xa13在叶片内的表达发生抑制作用,这一效应与XopN的毒性功能相关。相反,日本晴显性感病基因OsSWEET11/Xa13却受病菌侵染的诱导,在叶片内的表达水平大幅度提高。IRBB208和Asominori携带OsSWEET11/Xa13同源基因,该同源基因在叶片内的表达水平也因病菌侵染而大幅提高。在这4个水稻品种上,PXO99^A和∆XopN/XopN能够诱导OsSWEET11/Xa13或其同源基因表达,但∆XopN无此作用。另外,XopN对病菌在非寄主植物烟草上诱发过敏反应有量变贡献,相比PXO99^A或∆XopN/XopN,∆XopN引起过敏反应的程度有所降低。【结论】XopN是一个有限广谱性效应子,在拥有OsSWEET11同源基因的水稻品种上发挥毒性作用。XopN也是病菌繁殖所需要的,对病菌在非寄主植物上诱导过敏反应有一定贡献。