进境船舶食品舱小麦印度腥黑穗病菌

小麦印度腥黑穗病菌是一种世界性的检疫性的有害生物,我国迄今未有报道.     

进境面粉中印度腥黑穗病菌冬孢子的萌发试验

１９９７年南京局连续三次从进境船舶食品舱、进境邮寄物面粉中发现了印度腥黑穗病菌冬孢子,为了确定面粉中印腥冬孢子是否具有活力,我们进行了萌发试验,并用ＰＣＲ进行了验证。现将有关结果总结如下。１材料和方法１．１供试菌株供试菌株为印度腥黑穗病菌Ｔｉｌｅｔｉ...     

面粉中小麦印度腥黑穗病菌的检疫鉴定

面粉中小麦印度腥黑穗病菌检疫的关键要点是分离孢子.为此,本文就如何从面粉样品中简便、快捷地分离与获得干净冬孢子进行了研究,并根据获得孢子多少与难易情况,采用30个孢子直径统计法和套式PCR检测法.与常规方法相比,这2种方法可以使检测时间缩短4～10倍.在样品中孢子极少的情况下,采用套式PCR检测方法,仅2个孢子就能作出定性判断,可以有效地避免小麦印度腥黑穗病菌漏检,进而提高口岸疫情检出率.     

小麦印度腥黑穗病——我国小麦生产的又一潜在威胁

1931年以来,小麦印度腥黑穗病由一个在原发地为害轻微的地方性病害发展成为危及小麦生产、影响国际贸易的世界性检疫病害,再度显示出一种危险性病害对人类社会生活的重大冲击。70年代末期,我国开始关注小麦印腥病菌可能对我国小麦生产存在的潜在威胁,并进行了一些基础研究,1986年,我国公布此病为检疫性病害,迄今至少有40余个国家将此病列为检疫性病害,其中包括美国、加拿大、欧盟及前苏联各国,尤其是此病于1972年传人墨西哥后,几年之内使设立在墨西哥的国际小麦、玉米种质资源中心的一些小麦品系受到感染,因而此病有可能随同国际小麦种质资源的交换而加速扩散,1996年3月,美国农业部宣布在亚利桑那州发现此病,使我国     

小麦印度腥黑穗病菌的截获及各国检疫措施

本文就从美国面粉中截获小麦印度腥黑穗病菌的过程作了介绍，并综述了该病原菌与植物检疫的有关情况以及世界各国针对该病原菌采取的检疫措施。     

小麦印度腥黑穗病菌鉴定方法的新进展

自1930年印度西北部Haryana邦的Kamal地区首次发现小麦印度腥黑穗病(Tilletia indicaMitra,以下简称印腥)以来,由于感病寄主的广泛种植和适宜环境条件的存在,该病已在世界范围内的不同生态区迅速传播蔓延,在70年左右的时间内从一个危害轻微的地方性病害发展为威胁世界小麦生产和贸易的危险性真菌病害.     

一种简单快速鉴定面粉中孢子的方法

面粉常携带多种检疫性病虫,特别是易携带一类危险性病原菌小麦矮腥黑穗病菌TCK和小麦印度腥黑穗病菌TIM.因此,对面粉的检验主要是针对上述两种病原菌.2003年我们共检验面粉样品177份,占全局船舶检疫送检样品的52%,其中,截获TCK4批次,TIM10批次,较去年提高了250%.     

Test validation for the detection of Tilletia indica Mitra by multiplex real‐time PCR

Tilletia indica Mitra is a fungal pathogen causing Karnal bunt of wheat. Tilletia indica is a quarantine pest in many countries worldwide. In the European Union, imported wheat grain from countries where the fungus is present must be checked for the presence of T. indica teliospores. The inspection services at the borders need rapid, sensitive and reliable detection tests to identify T. indica spores on wheat grain. In this work, validation was carried out according to EPPO Standard PM 7/98 to evaluate the multiplex real‐time PCR test described in ISPM 27 Diagnostic Protocol for regulated pests (Annex 4 Tilletia indica Mitra) by means of a test performance study with nine participating laboratories, and the performance characteristics of the test were established. The original protocol was modified with regard to the extraction of DNA from the pellet obtained from the ‘washing test’ and the enrichment PCR step in order to increase the amount of template DNA for the real‐time PCR. The optimized test still has five teliospores as the limit of detection for the contaminated pellet but has an increased analytical sensitivity and had positive results with three teliospores in 93% of cases instead of 43% for the original test. The two closest Tilletia species, Tilletia horrida and Tilletia walkeri, were used to evaluate analytical specificity (exclusivity) and no cross‐reactions were obtained. Diagnostic sensitivity, diagnostic specificity, accordance and concordance were also evaluated.     

A simple bioassay for detecting and characterising insecticide resistance

A simple and rapid paralysis assay was developed to detect and characterise knockdown resistance in larvae of the house fly and pink bollworm. Pyrethroidtreated larvae unable to perform a stereotypic curling movement when probed with a warm needle, 10 min (house fly) or 60 min (pink bollworm) after treatment, were considered to be paralysed. Responses in a susceptible strain of house fly were compared with those of the pyrethroid-resistant strains kdr and super-kdr. In this assay, the kdr strain displayed over 28 fold resistance to deltamethrin, and super-kdr larvae were unaffected by doses up to 100μg. These results are in good qualitative agreement with previous studies. The assay detected no significant fenvalerate resistance in pink bollworm larvae collected from the field; this was consistent with mortality bioassays performed on wild adult males. The limitations and potential utility of the paralysis bioassay in resistance screening are discussed.     

一种简便易行的栗疫病菌单孢分离方法

介绍了一种分离栗疫病菌单分生孢子的简便易行的方法。此方法可以排除菌丝段的干扰,也适用于分生孢子器中产生微小分生孢子的其他真菌。     

一种用玉米拟轮枝镰孢穗腐病病粒制备接种体的简易方法

制备接种体是玉米穗腐病抗性鉴定中的一个重要环节, 本研究提出了一种用玉米拟轮枝镰孢穗腐病病粒制备接种体的简易方法, 即将上年的拟轮枝镰孢穗腐病带菌病粒按一定重量加入无菌水洗脱制成接种体。田间接种试验结果表明, 采用本方法简易制备的接种体接种后可获得与常规制备接种体相当的接种效果, 且在-18℃冻存15 d仍可达到新鲜的常规制备接种体接种后的水平。使用简易制备接种体对33个玉米品种进行了田间穗腐病抗性鉴定, 21个品种表现为感病或高感。与实验室常规培养制备接种体的方法相比, 本制备方法简便易操作, 用时短, 成本低, 无需专业仪器设备, 能够一次制备大量接种体, 可广泛用于玉米种质材料和新品种的拟轮枝镰孢穗腐病田间抗性鉴定及抗性材料筛选等。     

小麦矮化腥黑穗病菌冬孢子的定量检测及几种计数方法的比较

用人工污染有小麦矮化腥黑穗病菌Tilletia controversa Kuehn冬孢子的小麦样品，按国家标准GB／T18085－2000规定的洗涤程序，对TCK的定量检测以及3种冬孢子计数方法进行了比较研究，并建立了定量评估样品中孢子量的方法。研究表明，使用国际种子检验协会（ISTA）的标准种子洗涤方法仅能发现30．72％－46．58％的TCK孢子，检验洗涤后的小麦表面，仍可分析有大量孢子沾附于小麦种子表面。应用回归分析，建立了定量评估样品中孢子的方程Y＝2．0131（X＋2）^1.0325-2。通过比较3种孢子计数方法，认为依据国标建立的计数方法得到的结果变异最小。     

一种诊断昆虫微孢子虫病的简易方法

介绍一种利用鉴别寄主感染组织形态特征的变异来诊断昆虫微孢子虫病,鉴别微孢子虫种的方法。     

A simple method for a mini-preparation of fungal DNA

A simple method was established to prepare DNA from fungal mycelia cultured on an agar plate. The fungi tested successfully with this method contained Zygomycetes, Ascomycetes, Basidiomycetes, and Oomycetes. This method did not require any time-consuming steps to crush or digest mycelia or fractionation in a phenol–chloroform mixture. The DNA was easily extracted by immersing and dispersing the mycelial plugs in a specific buffer (200 mM Tris-HCl, 50 mM ethylenediaminetetraacetic acid, 200 mM NaCl, 1% n-lauroylsarcosine, pH 8.0), then concentrated by ethanol precipitation. The total time to complete the whole procedure was less than 1 h. The quality and quantity were sufficient for polymerase chain reaction amplification and Southern blot analysis.     

A rapid and simple method for determining fungicide resistance in Botrytis

A simple test based on the germination of conidia of Botrytis on agar media augmented with various fungicides has been developed. Average concentrations causing a 50% reduction of germ-tube growth (EC₅₀) of highly sensitive isolates were determined on 1% malt extract agar (thiophanate-methyl 0.090 ppm; iprodione 0.566 ppm; fludioxonil 0.026 ppm; fenhexamid 0.144 ppm), 1% malt extract agar with 100 ppm salicyl hydroxamic acid (QoI fungicides, viz. trifloxystrobin 0.009 ppm; pyraclostrobin 0.013 ppm; azoxystrobin 0.087 ppm), 0.5% yeast extract agar (boscalid 0.069 ppm) and 0.5% sucrose agar (cyprodinil 0.053 ppm). In order to detect different levels of resistance against these various fungicides, two discriminatory concentrations were identified for each compound. A routine assay method was developed in which drops of a conidial suspension harvested directly from diseased plant material or sporulating cultures were incubated on each of 20 different agar media. Because of a very short time-span of 24–48 h between sample collection and evaluation of results, field-specific information on the occurrence, frequency and types of resistance of Botrytis against common botryticides in soft-fruit production may be generated prior to the main fungicide spray season at blossom time.