紫茎泽兰对7种牧草种子萌发及幼苗生长的化感作用

检测了紫茎泽兰地上部分榨取液对7种牧草种子萌发和幼苗生长的化感作用。结果表明：（1）紫茎泽兰地上部水浸提液对7种供试牧草种子的萌发均表现为强烈的抑制作用（P 〈0.05）,对光叶紫花苕种子萌发的抑制作用显著小于对紫花苜蓿、白三叶、红三叶、东非狼尾草、非洲狗尾草和鸭茅种子萌发的抑制作用（P 〈0.05）;（2）从幼苗生长的测定结果分析,紫花苜蓿,红三叶和光叶紫花苕对低浓度紫茎泽兰叶片水提液不敏感,对高浓度提取液较敏感。白三叶、鸭茅、非洲狗尾草和东非狼尾草对紫茎泽兰的化感作用较敏感;（3）根据化感作用检测结果,分析认为光叶紫花苕、红三叶和紫花苜蓿是紫茎泽兰替代种植的适宜草种。     

基于双向电泳结合质谱分析燕麦种子萌发过程蛋白的变化

为分析燕麦(Avena sativa L.)种子蛋白质在萌发过程中的变化,探求燕麦种子吸胀萌发的机理。采用双向电泳结合基质辅助激光解析飞行时间质谱的方法对燕麦品种白燕2号种子吸涨萌发0 h和25 h的蛋白质进行了分离和鉴定。结果表明:燕麦种子蛋白在吸水萌发25 h后18个蛋白点的表达发生了显著变化;这些蛋白质点经质谱分析并搜索NCBI数据库,有10个蛋白点得到明确鉴定,其中7个蛋白质点表达量在燕麦种子萌发后下降,3个表达量增加。差异表达的蛋白质包括12s球蛋白、种子贮藏球蛋白和醇溶燕麦蛋白-3等蛋白。这些蛋白表达量的变化与燕麦种子萌发进程有关,为进一步从蛋白质水平来探索燕麦种子萌发机理提供了理论依据。     

施肥和刈割对紫茎泽兰和黑麦草苗期竞争的研究(简报)

通过种植牧草抑制外来入侵种紫茎泽兰是一条重要的替代控制途径。为探究施肥和黑麦草刈割等管理措施对紫茎泽兰与多年生黑麦草竞争的影响,运用de Wit取代系列设计,设置不施肥与施肥2个肥力水平、黑麦草不刈割与刈割2个干扰水平,对紫茎泽兰和黑麦草苗期的相对竞争表现进行了研究,同时观察了紫茎泽兰苗期的形态特征变化。结果表明,施肥和黑麦草刈割2个因素对紫茎泽兰的相对产量(RY)有显著互作效应,但对黑麦草RY没有互作效应。施肥能提高紫茎泽兰的相对竞争力,而刈割黑麦草可以降低紫茎泽兰的竞争力;在所有管理处理下,黑麦草的竞争强度均大于紫茎泽兰。紫茎泽兰的株高、分枝数和生物量在施肥和不刈割黑麦草处理下达到最大,而在不施肥和刈割黑麦草处理下最小。建议在与本试验地类似的土壤中,不施或少施肥并增加黑麦草的刈割次数,来降低紫茎泽兰的竞争力,以提高黑麦草的替代控制效果。     

紫茎泽兰在凉山金阳的分布情况及其规律

调查了紫茎泽兰(Eupatorium adenophorum Spreng)在金阳县的分布规律,为防止紫茎泽兰对金阳县农业生产和生态环境破坏提出了一些依据。本次调查了每个样方内的紫茎泽兰的植株数、平均高度、单株生物量和总生物量,发现紫茎泽兰的总株数从海拔1 650 m的3株增长到580 m的32 640株,极差达到32 637,变化极其显著;紫茎泽兰的平均高度和单株生物量变化也较大;同时发现紫茎泽兰的植株数、平均高度、单株生物量和总生物量随海拔的升高成负相关,其中单株生物量与海拔的相关性最显著,相关系数达到-0.937 4。     

紫茎泽兰在凉山金阳的分布情况及其规律

调查了紫茎泽兰（Eupatorium adenaphorum Spreng）在金阳县的分布规律，为防止紫茎泽兰对金阳县农业生产和生态环境破坏提出了一些依据．本次调查了每个样方内的紫茎泽兰的植株数、平均高度、单株生物量和总生物量，发现紫茎泽兰的总株数从海拔1650m的3株增长到580m的32640株，极差达到32637，变化极其显著；紫茎泽兰的平均高度和单株生物量变化也较大；同时发现紫茎泽兰的植株数、平均高度、单株生物量和总生物量随海拔的升高成负相关，其中单株生物量与海拔的相关性最显著，相关系数达到-0．9374．     

紫茎泽兰对16种牧草发芽及幼苗生长的化感作用

外来入侵植物紫茎泽兰目前广泛分布于我国西南地区,给当地草地畜牧业带来极大危害。牧草替代种植是紫茎泽兰严重危害地区草地生态恢复的有效途径。本研究采用生物试验方法,检测了紫茎泽兰地上部分5%的榨取液对我国亚热带种植的16种主要牧草,共38个品种的种子萌发和幼苗生长的化感作用。结果表明,紫茎泽兰对多数牧草发芽势有显著抑制作用(P<0.05),但只对少数牧草的发芽率、死亡率和败育率有显著影响(P<0.05),对所有幼苗生长都没有明显抑制作用;紫茎泽兰对不同草种的化感潜力存在较大差异,但对种内不同品种间的影响差异较小,对少数草种的化感作用不明显;紫茎泽兰对部分牧草幼苗生长具有显著促进作用(P<0.05)。根据化感作用检测结果,初步认为非洲狗尾草、鸭茅、紫花苜蓿、大翼豆、箭筈豌豆和光叶紫花苕是紫茎泽兰牧草替代种植的适宜草种。     

紫茎泽兰传播规律研究

紫茎泽兰(EupatoriumadenophorumSpreng)作为一种世界性的恶性杂草,近年来在四川地区蔓延迅速,通过查阅资料,结合调查、标本描述、选点试验等手段,对全省范围内紫茎泽兰进行了系统地研究,获得了紫茎泽兰的植物学特性、紫茎泽兰在四川的传入历史情况、传播途径、分布与蔓延的趋势和规律,以及紫茎泽兰对畜牧业、农业、林业的危害情况等资料。同时分析了气候、地理等自然因素对紫茎泽兰生长影响的规律,为对其进行有效防除和利用奠定了基础。     

九龙县紫茎泽兰分布、危害调查与对策

对紫茎泽兰的生境、分布范围、危害程度、生物学特性、生长繁殖规律进行了调查,掌握了紫茎泽兰的生物学特性和生长繁殖规律,对防治紫茎泽兰提出了初步对策,为彻底铲除紫茎泽兰提供了可靠的科学理论依据。     

紫茎泽兰处理鸡柔嫩艾美耳球虫后差异基因筛选和分析

为获得紫茎泽兰处理鸡柔嫩艾美耳球虫后差异基因,将0.5%紫茎泽兰提取液作用于鸡柔嫩艾美耳球虫卵囊孢子化过程,采用抑制消减杂交技术（SSH）筛选处理后卵囊的差异表达基因,通过GO和COG功能预测分析,并采用实时荧光定量PCR验证差异表达的基因。结果显示,采用SSH成功构建了紫茎泽兰处理前后鸡柔嫩艾美耳球虫卵囊的cDNA消减文库,获得86条ESTs序列,经拼接和聚类后得到31条独立基因（Unigenes）,其中有23个基因有功能注释,8个基因没有功能注释,另外有4个基因没有同源性匹配。为进一步验证文库的特异性,从中随机选取3个差异表达基因,运用实时荧光定量PCR技术验证表面抗原13、3-羟酰辅酶A脱氢酶和细胞色素P450基因在紫茎泽兰处理前后的表达差异,结果显示,3个基因在经紫茎泽兰处理的鸡球虫孢子化卵囊中的表达量明显低于未处理组,说明紫茎泽兰对柔嫩艾美耳球虫卵囊具有一定的活性抑制作用。本试验获取了紫茎泽兰作用柔嫩艾美耳球虫卵囊的主要调控基因,为将紫茎泽兰研发成环境杀虫剂奠定了良好的理论基础,也可为球虫致弱疫苗或基因缺失疫苗的靶标筛选研究提供参考。     

紫茎泽兰的生物学特点及其在畜牧生产中的应用

紫茎泽兰在我国分布范围广泛,是一种可利用的资源。目前有大量研究在探讨紫茎泽兰的生物学功能和提取其有效的活性成分,可以通过资源化利用提高紫茎泽兰的经济价值,变废为宝。论文综述了紫茎泽兰的生物学特点、生物学活性及其在动物抗寄生虫、抑制细菌、免疫调节和作为饲料原料的应用研究现状,为后期紫茎泽兰的研究提供参考。     

紫茎泽兰浸提液对牧草种子发芽和幼苗生长的影响

紫茎泽兰被我国列为16种有害外侵物种之首,在西南地区大规模侵入草场、农田、森林,很有必要了解其化感效应,研发无害化处理与资源化利用相结合的技术。试验以广泛分布于四川省凉山州的白三叶、黑麦草和紫花苜蓿为材料,比较了新鲜(extract of fresh E. adenophorum, EFA)和腐熟紫茎泽兰的浸提液(extract of composed E. adenophorum, ECA)对牧草种子萌发和幼苗生长的影响,以明确这种外侵植物对草场植被的危害作用和腐熟处理效果。随EFA浸种和培养浓度的提高,不同程度地抑制牧草种子发芽和幼苗生长。当浓度达到100 mg/L EFA时,牧草根毛消失,根尖向上卷曲,离开EFA,根尖发黑,甚至死亡。相反,用铜绿假单胞菌(Pseudomonas putita sp.)和高温纤维菌(Clostridium thermocellum sp.)组成的混合菌剂腐熟紫茎泽兰后,ECA提高牧草种子发芽率、发芽指数和活力指数,并促进幼苗生长,其最大增幅达到21.11%(种子发芽率),24.12%(苗高)和22.48%(生物量)。此外,用100 mg/L EFA浸种和培养牧草幼苗,抑制胚乳中的淀粉、蛋白质和肌醇磷酸盐水解,显著降低游离氨基酸、可溶性糖、可溶性磷,以及幼苗中的叶绿素、根系活力及硝酸还原酶活性,ECA则相反。这可能是EFA抑制(或ECA促进)牧草种子发芽和幼苗生长的重要生理原因之一。因此,EFA含有对牧草有害化感的物质,抑制其种子发芽和幼苗生长;腐熟紫茎泽兰可降解毒素,刺激牧草种子发芽,促进幼苗生长,实现紫茎泽兰的无害化处理与资源化利用,为当地农业和畜牧业生产提供大量的优质有机肥源。     

家蚕雌性附腺分泌蛋白的双向电泳分离及质谱鉴定

采用24 cm的双向电泳可以分离出家蚕雌性附腺分泌的70多种蛋白或多肽,其中有30多个蛋白或多肽的表达量较大。从这些蛋白斑点在凝胶中的分布位置上看,大多数分布在等电点pI 4~7的范围内,主要蛋白的分子量较大。对其中表达量相对较高的33个蛋白点进行胶内酶解后质谱分析,并根据检索到的蛋白质的功能进行分类,最多的是核糖体蛋白家族,占蛋白质总量的30.4%,另外还有2个热休克蛋白。除此之外,还有一些如RNA结合蛋白、转录延长因子等功能性蛋白质。这些蛋白质共同组成了具有特殊物理性质的分泌物。     

牛源大肠杆菌不同生长阶段的比较蛋白质组学研究

为了分析牛源大肠杆菌生长周期中不同阶段蛋白的表达变化,本研究采用二维凝胶电泳结合质谱技术分离并鉴定了大肠杆菌生长周期中延迟期、对数期和平稳期菌体全蛋白的表达变化。结果显示,大肠杆菌菌体全蛋白组成在延迟期、对数期和平稳期存在差异,大肠杆菌生长延迟期的外膜蛋白W和磷酸丙酮酸羧化酶表达量高于对数期和平稳期,而周质蛋白、外膜蛋白Ⅱ和磷酸转乙酰酶表达量低于对数期和平稳期。本研究结果初步揭示了菌体蛋白表达变化与细菌生长的关系,为进一步用于大肠杆菌奶牛乳房炎的防控提供参考。     

Identification of abundant proteins and potential allergens in Culicoides nubeculosus salivary glands

IgE-mediated type 1 hypersensitivity reactions to the bites of insects are a common cause of skin disease in horses. Insect bite hypersensitivity (IBH) is most frequently associated with bites of Culicoides spp. and occurs in all parts of the world where horses and Culicoides coexist. The main allergens that cause IBH are probably some of the abundant proteins in the saliva of Culicoides associated with blood feeding. Western blots of Culicoides proteins separated by 1D gel-electrophoresis detected strong IgE responses in all horses with IBH to antigens in protein extracts from wild caught Culicoides, but only weak responses to salivary antigens from captive bred C. nubeculosus which may reflect important differences among allergens from different species of Culicoides or differences between thorax and salivary gland antigens. 2D electrophoresis and mass spectrometry were used to identify several of the abundant proteins in the saliva of C. nubeculosus. These included maltase, members of the D7 family, and several small, basic proteins associated with blood feeding. The most frequently detected IgE-binding proteins were in a group of proteins with pI>8.5 and mass 40-50kDa. Mass spectrometry identified two of these allergenic proteins as similar to hyaluronidase and a heavily glycosylated protein of unknown function that have previously been identified in salivary glands of C. sonorensis.     

Effect of condensed tannin ingestion in sheep and goat parotid saliva proteome

Saliva appears as a defence mechanism, against potential negative effects of tannins, in some species of animals which have to deal with these plant secondary metabolites in their regular diets. This study was carried out to investigate changes in parotid saliva protein profiles of sheep (Ovis aries) and goats (Capra hircus), induced by condensed tannin ingestion. Five Merino sheep and five Serpentina goats were maintained on a quebracho tannin enriched diet for 10 days. Saliva was collected through catheters inserted on parotid ducts and salivary proteins were separated by two-dimensional gel electrophoresis. Matrix-assisted Laser desorption ionization - time of flight (MALDI-TOF) and liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to identify the proteins whose expression levels changed after tannin consumption. Although no new proteins appeared, quebracho tannin consumption increased saliva total protein concentration and produced changes in the proteome of both species. While some proteins were similarly altered in both species parotid salivary protein profile, sheep and goats also presented species-specific differences in response to tannin consumption.     

二化性家蚕卵低温和常温催青的胚胎期蛋白质表达差异分析

家蚕二化性品种的滞育性受上代胚胎期环境条件调控,查找家蚕胚胎期滞育关联蛋白,可为最终阐明家蚕滞育的分子机制提供实验依据。以家蚕二化性品种秋丰的蚕卵为材料,分别在25℃常温和18℃低温条件下催青,提取胚胎不同发育时期蚕卵的易溶性和难溶性蛋白,采用蛋白质双向电泳(2-DE)和图像分析技术,研究蚕卵在胚胎不同发育时期的蛋白质差异表达谱。在易溶性和难溶性蛋白图谱中分别检测到5个和7个差异显著的蛋白点。对这些差异蛋白点进行质谱鉴定,有8个蛋白点得到可信的最佳匹配蛋白报告,共鉴定出卵特异蛋白、卵黄原蛋白、表皮蛋白和丙酮酸脱氢酶激酶4种蛋白。这些差异表达蛋白可能导致蚕卵胚胎中物质代谢和能量代谢的不同,据此推测低温和常温催青可能在蚕卵胚胎中诱导了不同的生理生化反应。     

Evaluation of differentially expressed proteins following serum exposure in avian pathogenic Escherichia coli

Colibacillosis, caused by avian pathogenic Escherichia coli (APEC), is an extraintestinal disease that causes great economic loss to the poultry industry each year. APEC must overcome host defenses, such as immune system components found in serum, in order to establish infection; however, the mechanism of such serum resistance has been elusive. In the present study, a proteomic approach was used to evaluate APEC proteins that were differentially expressed after exposure to chicken serum to identify specific proteins that may be involved in serum resistance of APEC isolates. Proteins were isolated and separated by two-dimensional (2D) gel electrophoresis, and 10 protein spots corresponding to differentially expressed proteins were chosen for sequencing using electrospray ionization tandem mass spectrometry. Eight proteins were identified among the spots, some of which have previously been associated with the virulence of E. coli. Significantly, an outer-membrane protein previously associated with serum resistance, OmpA, was among those proteins identified, further indicating that differential regulation of this protein may be involved in serum resistance. This study opens the door to future research using a proteomic approach to identify the key players in serum resistance of APEC.     

Host cell-specific protein expression in vitro in Ehrlichia ruminantium

Ehrlichia ruminantium, a tick-transmitted pathogen, is the causative agent of heartwater in ruminants. In this study, a proteomic approach was used to identify host cell-specific E. ruminantium proteins encoded by the map1 multigene family, expressed in vitro in bovine endothelial and tick cell cultures. Two-dimensional gel electrophoresis combined with mass spectrometry analysis was used to establish the identities of immunodominant proteins. Proteins extracted from E. ruminantium-infected endothelial cells were shown to be products of the map1 gene, whereas tick cell-derived E. ruminantium proteins were products of a different gene, map1-1. The expressed proteins were found to be glycosylated. Differential expression of MAP1 family proteins in vitro in mammalian and tick cell cultures indicates that the map1 multigene family might be involved in the adaptation of E. ruminantium to the mammalian host and vector tick.     

Proteomic survey of the pathogenic Mycoplasma hyopneumoniae strain 7448 and identification of novel post-translationally modified and antigenic proteins

Mycoplasma hyopneumoniae is an important pathogen for pigs, being the causative agent of enzootic pneumonia. Recently, the genome sequences of three strains, J, 7448 and 232 have been reported. Here, we describe the results of a proteomic analysis, based on two-dimensional gel electrophoresis of soluble protein extracts, immunoblot and mass spectrometry, which was carried out aiming the identification of gene products and antigenic proteins from the M. hyopneumoniae pathogenic strain 7448. A preliminary M. hyopneumoniae proteome map in two pH ranges (3-10 and 4-7) was produced. A total of 31 different coding DNA sequences (CDSs), including three hypothetical ones, were experimentally verified with the identification of the corresponding protein products by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. According to the Clusters of Orthologous Groups (COG) functional classification, the identified proteins were assigned to the groups of metabolism (13), cellular processes (5) and information and storage processing (4). Nine of the identified proteins were not classifiable by COG, including some related to cytoadherence and possibly involved in pathogenicity. Moreover, at least five highly antigenic proteins of M. hyopneumoniae were identified by immunoblots, including four novel ones (a heat shock protein 70, an elongation factor Tu, a pyruvate dehydrogenase E1-beta subunit and the P76 membrane protein). The now available proteome map is expected to serve as a reference for comparative analyses between M. hyopneumoniae pathogenic and non-pathogenic strains, and for methabolic studies based on cells cultured under modified conditions.