利用BIOLOG鉴定系统快速鉴定菜豆萎蔫病菌的研究

 本研究利用美国Biolog公司生产的MicroStation^TM V3.5系统对我国一类危险性病害菜豆萎蔫病菌及其相关菌进行了快速鉴定研究。研究结果表明,来自不同国家和寄主的24株菜豆萎蔫病菌及其相关致病变种,23株准确鉴定至种水平,其中13株鉴定至致病变种水平,种水平的鉴定准确率为95.8%,另外1株至属水平。同时2株苜蓿萎蔫病菌和2株番茄溃疡病菌均鉴定至致病变种水平。经聚类分析研究,结果支持了对格兰氏阳性植病细菌在属、种水平的分类。本研究是首次使用该系统对格兰氏阳性植病细菌进行鉴定研究,试验证明Biolog鉴定系统用于快速鉴定菜豆萎蔫病菌是一个很有用的工具,且由于其标准化程度高、快速准确,符合我国口岸植物检疫的要求。     

基于Brn1基因的大蒜紫斑病菌PCR检测技术

大蒜紫斑病菌是一种严重危害大蒜生产的真菌性病害,是中国大蒜出口中需要检疫的一种病害。根据1,3,8-三羟基萘还原酶基因(Brn1)的部分序列,设计出一对大蒜紫斑病菌特异性引物AP4/CTU,该引物特异性好,能专一扩增出480 bp电泳条带,而供试大蒜上的其他病害、不同属的真菌及空白对照均无扩增条带,建立了大蒜紫斑病菌的PCR鉴定方法。该方法灵敏度较高,DNA检测灵敏度为4 ng。该方法快速简便,适用于出入境检验检疫及大蒜健康检测领域。     

一种快速鉴定玉米细菌性枯萎病菌的新技术——噬菌体株系鉴别法

利用８株玉米细菌性枯萎病菌噬菌体专化性的差异，结合氯化三苯基四氮唑（２，３，５－ｔｒｉｐｈｅｎｙｌｔｅｔｒａｚｏｌｉｕｍｃｈｌｏｒｉｄｅ）对不同致病力细菌显色反应的不同，组成了一种新的细菌快速鉴定技术，称为噬菌体株系鉴别法，可鉴定玉米细菌性枯萎病菌直至株系。用这个方法鉴定了从美国、日本、南斯拉夫、西德和罗马尼亚进口的玉米种子中截获的许多玉米细菌性枯萎病菌菌株，结果准确快速，并和血清学鉴定及致病性测定的结果相一致     

油菜茎基溃疡病菌LAMP-LFD检测方法的建立

本文基于环介导等温扩增技术与横向流动试纸条相结合的方法,建立了一种应用于油菜茎基溃疡病菌(Leptosphaeria maculans)的LAMP-LFD快速检测方法。以油菜茎基溃疡病菌的ITS基因序列为靶序列,设计出一套用于LAMP-LFD检测的引物和探针,优化了反应体系与反应条件(63℃,35 min)。结果表明:只有油菜茎基溃疡病菌出现阳性条带,其他参照菌株和阴性对照均未出现阳性条带,说明LAMP-LFD检测特异性强;灵敏度检测表明,对油菜茎基溃疡病菌的检测极限可低至114 fg/μL,灵敏度比传统PCR高10倍;该方法可从进境船载油菜籽样品中成功检测出油菜茎基溃疡病菌,检测结果与传统的鉴定方法一致。LAMP-LFD检测方法能够快速检测油菜茎基溃疡病菌,具有简便、灵敏、特异性高,不依赖特殊检测设备等优点,极具推广前景。     

江西省蔬菜根肿病菌致病性分化研究

正芸薹根肿病菌(Plasmodiophora brassicae)存在生理小种分化现象,国际上通常采用Williams鉴别系统~([1])和欧洲根肿病菌鉴别寄主(ECD)系统~([2])鉴定根肿病菌生理小种。我国尚没有研究出适合本国根肿病菌生理小种鉴定的鉴别寄主系统。国内学者研究根肿病菌生理小种通常采用Williams鉴别系统~([3])。为了明确江西省蔬菜根肿病菌致病性分化及病菌小种的变化趋势,作者采用     

小麦抗根腐病突变体的获得及其鉴定

 本研究利用细胞工程技术将根腐病菌分生孢子定量直接接种在筛选出的小麦抗根腐病菌培养滤液的植株上进行连续抗病性鉴定,并在根腐病严重发病地区进行多年多点的抗病性鉴定,并在愈伤组织阶段和再生植株的幼苗阶段进行早期鉴定。     

两种向日葵检疫性真菌病害的多重DPO-PCR检测方法

本研究根据向日葵白锈病菌大亚基核糖体RNA基因序列,向日葵黑茎病菌的ITS-5.8S r RNA基因序列,分别设计特异性DPO(dual priming oligonucleotide)引物,建立同时检测这两种检疫性病菌的多重DPO-PCR检测方法,并对其特异性和灵敏度进行评价。结果表明,所设计的DPO引物特异性强,仅向日葵白锈病菌和向日葵黑茎病菌可分别扩增出307 bp与388 bp的特异性条带,其他参照菌株及阴性对照均无条带;检测体系对混合模板中向日葵白锈病菌和向日葵黑茎病菌的DNA灵敏度均达0.05 ng/μL;且该检测方法对退火温度不敏感,适用范围广。该方法能够准确、快速的检测向日葵白锈病菌和向日葵黑茎病菌,适合于口岸实验室的快速检测。     

木薯细菌性萎蔫病菌的检疫方法研究

本文对木薯细菌性萎蔫病菌的致病性测定、细菌的分离、细菌的培养条件和培养基选择、细菌的生理生化测定、分子生物学鉴定方法等方面进行了系统研究，确定了该病菌的菌落鉴定特征，建立了从木薯繁殖材料上进行病原菌检测的快速、灵敏、准确的PCR检测方法。     

甘蓝枯萎病菌1号和2号生理小种的快速检测与鉴定

 甘蓝枯萎病菌生理小种传统鉴定方法费时费力,不能满足生产的要求,因此需要建立一种快速、可靠的分子检测技术。本研究在甘蓝枯萎病菌1号和2号生理小种基因组测序的基础上,通过比较基因组学方法筛选1、2号生理小种各自的特异基因片段并设计引物,并分别以10个甘蓝枯萎病菌1号生理小种菌株、2个2号生理小种菌株、7个尖孢镰刀菌其他专化型菌株及4个外围菌株DNA为模板进行常规PCR扩增,筛选出甘蓝枯萎病菌1号和2号生理小种特异性引物,同时引入尖孢镰刀菌通用引物W106R/W106S,建立起一步三重PCR检测甘蓝枯萎病菌1、2号生理小种的分子检测技术。结果表明,该分子检测技术实现了在一次PCR反应中快速、准确地同步检测出甘蓝枯萎病菌DNA、罹病甘蓝组织和土壤中的甘蓝枯萎病菌1号和2号生理小种,对检测甘蓝植株是否感染枯萎菌及甘蓝种植区土壤是否受到枯萎菌的污染有实用价值。     

杨树角斑病病原菌分离及其培养特性研究

对杨树角斑病菌进行分离和鉴定,并研究不同培养条件对其培养特性的影响,以获得病原菌的纯培养并筛选出最佳培养条件,为杨树角斑病防治提供理论参考。分别采用组织和单孢分离法分离病原菌,根据子实体形态和病菌rDNA ITS序列对其进行鉴定。观察不同培养基、温度、光照、pH对病原菌生长的影响。结果表明,自然条件下,用杨树角斑病菌的分生孢子分离较病组织分离易成功。根据病菌的形态特征和rDNA ITS序列分析,将病菌鉴定为柳假尾孢[Pseudocercospora salicina(Ell.&Ev.)Deighton]。培养基、温度、光照、pH等因素影响病菌在培养基上的生长。燕麦培养基、pH6～7、25℃、黑暗条件是适合的培养条件。     

拟除虫菊酯类农药的免疫毒性研究进展

拟除虫菊酯类杀虫剂因其高效、对哺乳动物低毒和易降解等特性已被广泛使用,目前对其生态毒理学的研究主要集中在神经及内分泌干扰、发育和生殖毒性方面,而关于其免疫毒性的研究也越来越受到了关注。文章对拟除虫菊酯类农药的免疫毒性及其分子机制研究进展进行了总结,主要从免疫器官、免疫细胞、免疫分子和免疫功能等方面综述了此类杀虫剂对不同生物的免疫毒性及其可能的分子机制,同时总结了拟除虫菊酯类杀虫剂暴露与免疫相关疾病发生的关联,以期为拟除虫菊酯类农药免疫毒性的进一步探究提供参考。     

螺虫乙酯在染毒雄性大鼠体内的分布与代谢

建立了采用超高效液相色谱-串联质谱仪(UPLC-MS/MS)检测不同动物样品中螺虫乙酯及其主要代谢物残留量的方法,并研究了螺虫乙酯在大鼠体内的吸收与代谢。样品中螺虫乙酯及其主要代谢物经甲醇提取及C18固相萃取(SPE)柱净化后,用UPLC-MS/MS检测。结果显示,所建立方法快速、灵敏,每个样品上机检测仅需3 min,方法的最低检出浓度(LOQ)为0.005 mg/kg。对按照每千克体重每日250 mg剂量染毒28 d后的大鼠体内螺虫乙酯残留量的检测结果表明:螺 虫乙酯在睾丸、肝脏、肺、肾、心脏、血浆等器官和组织中的残留量较低,平均在0.012~0.025 mg/kg 之间,且分布差异不显著,而脂肪和肌肉中螺虫乙酯的残留量显著低于睾丸和肝脏中的残留量(P P 肾>血浆>肺>心脏>睾丸>脂肪>肌肉。     

Establishing targeted control of creeping perennial weeds with soil‐active chemical injections: Assessment of subterranean bud responses in contact

Increased infestation of aggressive creeping perennial weeds is a significant problem in urban vegetation management programs. These weeds produce vigorous biomass and extensive underground networks of either rhizomes or creeping roots that easily regrow from numerous buds. Foliar application of proper systemic herbicides has been a most effective way to suppress regrowth from underground creeping organs; however, killing the mature plants has disadvantages from aesthetic, economic, and ecological viewpoints. Therefore, we intended to test the possibility of soil‐injection of soil‐active herbicides for effective control of the perennial weeds which develop underground network systems. A pot experiment using combinations of seven species (seven rhizomatous and two having creeping roots) and five chemicals (four herbicides and a plant growth regulator) was conducted to assess whether and how chemicals diffused in soil affect the sprout and growth of buds on creeping organs. All the tested herbicides completely inhibited bud sprouting in one and more species when applied at median or high rates, while most of the flurprimidol‐applied segments sprouted but shoot elongation was significantly reduced. Characteristics of each herbicide were also reflected in the selectivity and features of new outgrowth. The results indicated that chemicals existing in soil were undoubtedly absorbed and affected bud activities. It is concluded that soil injection that delivers the probable soil‐active chemicals to subterranean creeping systems could be a promising technology for controlling noxious creeping perennials.     

塔克拉玛干沙漠腹地10种灌木光合器官的滞尘能力评价

通过全株光合器官取样,利用质量差减法,对塔中植物园10种灌木的滞尘能力进行多指标比较。结果表明：① 植物光合器官滞尘量与所处环境条件有关,在沙漠地区同种植物光合器官滞尘量大于沙漠以外区域。② 梭梭和沙拐枣的光合器官退化为同化枝,形态类似针叶树种的针叶,但尚有极度退化的叶片,其滞尘量大于其他区域针叶树种;与沙拐枣相比,梭梭同化枝节间距较短,叶片数量相对较多,因此梭梭的滞尘能力大于沙拐枣。③ 柽柳属植物虽然叶片也极度退化,但具有数量众多、分布密集、被覆绒毛、分泌盐分等特性,而且树冠光合器官密度大,具有很强的滞尘能力,光合器官滞尘量大于梭梭和沙拐枣,也大于阔叶灌木和阔叶乔木;由于光合器官表面滞尘特性、叶片数量以及树冠光合器官密度的差异,柽柳属植物光合器官滞尘能力也存在种间差异。④ 全植株光合器官滞尘量、单位重量光合器官滞尘量、单位表面积光合器官滞尘量都只能从一个侧面反映植物光合器官的滞尘能力,而单位树冠体积的光合器官滞尘量则可较全面地反映植物光合器官的滞尘能力。以单位体积树冠空间光合器官滞尘量为评价指标,10种灌木光合器官滞尘能力的排序为：甘蒙柽柳>长穗柽柳>华北柽柳>紫杆柽柳>刚毛柽柳>多花柽柳>多枝柽柳>沙生柽柳>梭梭>头状沙拐枣。     

羊草发育过程中同工酶的研究

采用聚丙烯酰胺凝胶电泳法 ,对羊草不同物候期不同器官的酯酶和过氧化物酶同工酶的多态性研究结果表明 :羊草酯酶同工酶活性在地上与地下营养器官间存在明显的互补性 ;过氧化物酶活性与有性生殖器官的发育有一定的一致性。总的来看 ,过氧化物酶同工酶与羊草的生长发育的关系较酯酶密切 ,可以作为生长发育特性评价较好生理指标 ;酯酶的稳定性较强 ,标志性酶带较恒定 ,可以作为遗传鉴定的指标。     

Histochemical demonstration of five enzymes' activities in Macrogyrodactylus clarii (Monogenea: Gyrodactylidae) from the catfish Clarias gariepinus

Histochemical techniques were applied to whole mounts, to study the distribution of the enzymes alkaline phosphatase, acid phosphatase, adenosine triphosphatase, 5'-nucleotidase and glucose-6-phosphatase in the organs and tissues of a viviparous monogenean, Macrogyrodactylus clarii Gussev, 1961, from the gills of the North African catfish Clarias gariepinus (Burchell) in Egypt. The following organs and tissues were studied: head region, anterior adhesive glands, mouth region, pharynx, intestine, testis, vesicula seminalis, male accessory gland, male accessory reservoir, copulatory organ, receptaculum seminis, egg-cell forming region, embryonic cells, excretory system, nerve cells, haptor, muscle fibres and subtegumental cell bodies (cytons). The enzymes showed marked differences in their activities among the studied organs and tissues. Alkaline phosphatase and acid phosphatase activities were detected in many organs and tissues, while the activities of adenosine triphosphatase, 5'-nucleotidase and glucose-6-phosphatase were restricted to a few organs. Although no positive reaction for any enzyme was observed in the anterior adhesive gland cells, a positive reaction for acid phosphatase was detected in the anterior adhesive areas. All enzymes showed marked activity in the digestive and excretory systems. The distribution of the enzymes in the tissues and organs of M clarii is compared with those of other monogeneans, including other gyrodactylids parasitizing the same host fish. Some possible functions of the enzymes are discussed.     

Sense organs of the nematodes Trichinella pseudospiralis Garkavi, 1972 and T. nativa Britov et Boev, 1972

The structure and distribution of the sense organs of the head and the tail of two day-old Trichinella species (females) have been determined by transmission (TEM) and scanning (SEM) electron microscopy. In both species occur 16 cephalic sense organs, which contain three modified dendritic processes, in addition to the amphids. The nerve cell bodies are situated in the area above the nerve ring. The amphid contains ten dendritic processes. The sense organs open to the external environment by pores through the cuticle. Above the bulbous tip, the dendritic processes are surrounded by hypodermal cells. They enter posteriorly the pseudocoelom and join in subventral, subdorsal and two sublateral nerves, with transparent nuclei, in the area above the nerve ring. The distribution of the sense organs of larvae from the muscles and from the uterus is similar to that of the adults. The dendritic processes are short, most of their distal chamber is compacted with a dense, filamentous material. The rectum in the tail end of the female is innervated from the dorsal ganglion composed by multipolar cells with numerous dendrites and a single axon. Close to the excretory pore occurs a hemizonid which consists of six to eight dendrites and is situated between the hypodermis and the muscles. This organ has not been found in the larva.     

毛缺沟姬蜂雌性内生殖器官的结构及其发育

报道了毛缺沟姬蜂Lissonota setosa(Geoffroy)成虫雌性内生殖器官的结构及其发育的系统观察结果。雌性成虫内生殖器官包括一对卵巢、两条侧输卵管、一条中输卵管、一个受精囊和一个附腺。成虫期卵巢生长速度随日龄是先加快后变慢。成虫期内其它生殖器官一般随日龄的增长而增大,达到一定程度后基本保持稳定。     

Review of the sequential development of Loma salmonae (Microsporidia) based on experimental infections of rainbow trout (Oncorhynchus mykiss) and Chinook salmon (O. tshawytscha)

Loma salmonae is a common gill parasite of salmonids, and essentially all species in the genus Oncorhynchus are susceptible. Infections occur in both fresh and salt water. Loma salmonae is directly transmissible by ingestion of spores or infected tissue. The parasite infects the wall of blood vessels of various organs, but the gill is the primary site of infection. Initial infection occurs in the intestine, and xenomas are easily detected in the gills by standard histology at 4-6 wk post-exposure. A few presporogonic stages of the parasite are found in the heart endothelium prior to xenoma formation in the gills. Ultrastructure studies of early infections demonstrated that wandering blood cells transport the meronts to the gills, and that merogony occurs in pillar cells and other cells underlying the gill endothelium. Xenomas develop in these cells, resulting in hypertrophied host cells filled with spores. Xenomas ultimately rupture, and are associated with severe inflammation in which free spores are found in macrophages. The parasites are most pathogenic during this phase of the infection, resulting in severe vasculitis and clinical disease. Both rainbow trout (Oncorhynchus mykiss) and Chinook salmon (Oncorhynchus ishawytscha) recover from infections, but free spores persist in kidney and spleen phagocytes for many months after xenomas are absent in Chinook salmon. Fish that have recovered from the infection show strong immunity against the parasite, lasting up to 1 year. Fish are susceptible to infection by other routes of exposure by spores; co-habitation, anal gavage, and intramuscular, intraperitoneal and intravascular injection. Autoinfection probably occurs following release of spores in blood vessels after xenomas rupture. The optimal temperature for L. salmonae infections is 15-17 degrees C, with a permissive range of 11-20 degrees C.     

棉花打顶对激素的影响与养分吸收变化

:研究棉花打顶后倒四叶内源激素和养分的关系表明:棉花打顶后,植株叶片的激素平衡关系被破坏,ABA/IAA和ABA/CTK比值上升,CTK含量下降.比值的变化降低了叶片中N和P的含量,但对K的变化无明显影响;降低了茎中N的含量,但对P和K的变化无明显影响;增加了生殖器官中的N,但对P,K的变化无明显影响.涂抹NAA能使叶片和茎中N下降的幅度缓解或加强.打顶对叶片的衰老影响最为明显.它将使棉株整体(尤其是营养器官)由正常生长向衰老方向转化,叶片内氮素营养减少,叶绿素含量下降.