大豆疫霉根腐病菌毒素产生条件的研究

 大豆疫霉根腐病(Phytophthora sojae)是大豆的重要病害之一,危害大,发展快。自1918年首次在美国印弟安那州发现此病以来、相继在澳大利亚、加拿大,匈牙利等国发生。     

大豆疫霉根腐病菌生理小种鉴定及毒性分析

对采自黑龙江省、吉林省的42株大豆疫霉根腐病茵进行生理小种鉴定，其中11株可被分别鉴定为1号、3号和8号生理小种。1号小种为优势小种，3号、8号小种为国内首次报道。其余31株在鉴别寄主上出现中间类型反应无法鉴定小种，但可以划分为12个毒性类型，病原茵群体具有很高的毒性多态性，其中有些菌株毒性较强。     

雪松疫霉根腐病菌

雪松疫霉根腐病菌(Phytophthora lateralis Tucker et Milbrath)是我国进境植物检疫性有害生物,可引起寄主植物严重的根腐病。鉴于该病原菌潜在的危险性,本文就其分布、寄主范围、传播方式、危害症状、形态特征以及检疫鉴定方法等进行了综述,供检疫鉴定参考。     

大豆疫霉根腐病菌游动孢子的产生因素

 本文对影响大豆疫霉根腐病菌游动孢子的产生因素进行了研究。8 d的菌丝体经蒸馏水浸泡,在25℃下培养72 h可以获得高浓度的游动孢子悬浮液,菌龄越短游动孢子浓度越大,偏酸性的环境有利于游动孢子产生,温度是影响游动孢子形成的重要因素,在0、35℃下均无游动孢子产生,游动孢子产生的适宜温度范围是20~30℃,最佳温度为25℃。     

安徽省大豆疫霉根腐病菌的鉴定及rDNA-ITS序列分析

为明确安徽省夏大豆疫霉根腐病的病原菌种类,对采集自涡阳、怀远、固镇3个县的夏大豆病株及土样分离纯化后获得28株菌株,选取6株代表性菌株,通过形态学观察及核糖体DNA-ITS序列分析对其进行鉴定,并测定了其致病型。结果表明,6株菌株在利马豆培养基上菌落白色,质地均匀;菌丝无隔,致密,具近直角分枝;在10%V8C培养液中,游动孢子囊顶生,不脱落,卵形至椭圆形,无明显乳突,有内层出现象,长宽比大于1.6∶1;同宗配合,在利马豆培养基上单株培养产生大量卵孢子,藏卵器球形,雄器大多侧生;接种合丰35大豆品种后出现典型的大豆疫霉根腐病症状。r DNA-ITS序列分析表明,6株菌株与Gen Bank中大豆疫霉Phytophthora sojae的ITS序列同源性高达100%;菌株GY4、GY8、HY11、HY16、GZ10、GZ21的毒力公式分别为1b,2,3a,3b,4,5,6,7;1b,1d,3a,3b;1d,3a,3b,3c,4,5,6,7;2,3c,4,5,6,7;1b,3a,3c,5,8;3a,3b,5,6,7,8;属于6个不同的致病型。研究表明,这6株菌株均为大豆疫霉。     

大豆疫霉菌检测技术研究进展

由大豆疫霉菌（Phytophthora sojae Kaufmann ＆ Gerdemann）侵染大豆引起的大豆疫霉根腐病（Phytophthora Root Rot of Soybean）是大豆生产上的毁灭性病害之一。该病害于1948年在美国印第安纳州首次发现，1955年公开报道后，日本、澳大利亚、新西兰、印度、加拿大、巴西、阿根廷、前苏联、匈牙利、德国、英国、法国、意大利、瑞士、埃及、尼日利亚等国家相继有了报道。     

大豆疫霉根腐病菌单游动孢子的毒性遗传与变异

 采用离体叶柄伤口接种法测定大豆疫霉根腐病菌44号生理小种单游动孢子连续3代或4代分离后代的毒性,结果表明:从S1中选择与亲本相比毒性不发生变异的1号单游动孢子菌株(44号生理小种)和变异最大的30号单游动孢子菌株(1号生理小种)继续分离2代或3代,单游动孢子毒性变异趋势主要是从44号生理小种变异为3号生理小种,也有变异成毒性公式为7或1a,7的单游动孢子。大豆疫霉根腐病菌无性世代毒性变异几率很高,多数单游动孢子毒性在分离后代中都发生变异,产生不同的小种或毒性公式,并且毒性变异基本不能稳定遗传。     

大豆疫霉病菌的分离及其生物学特性的研究

大豆疫霉病菌是我国重要的对外检疫对象之一,而栽培大豆又起源干我国。依据植物病理学中寄主与病原菌在原产地长期共存共同进化的原理,在我国极有可能有大豆疫霉病菌的存在,只是因为分离检测技术以及生态等方面的原因,较长时间内一直未能在我国分离到此病原菌。1989～1990年我们从大豆病组织中分     

黑龙江省大豆疫霉根腐病调查与病原分离

１９９６年对黑龙江省东部和中部大豆产区２３个市、县的大豆苗期疫霉根腐病进行了调查、研究,应用ＰＢＮＩＣ疫霉选择性培养基分离大豆疫霉根腐病病原菌,从牡丹江、穆棱、林口和佳木斯豆田具疫霉根腐症状的大豆植株上分离到大豆疫霉根腐病菌,并从根腐病株上单独或与大豆疫霉菌同时分离到终极腐霉菌,研究进一步证实我国黑龙江省有大豆疫霉根腐病。调查发现,大豆疫霉根腐病和终极腐霉根腐病主要发生在土质粘重、土壤含水量高或易积水的田块。     

大豆疫霉病菌在中国的发现及其生物学特性的研究

 本文通过对从黑龙江、吉林和北京等地的大豆上分离的7个疫霉菌株的一系列生物学特性的研究,并以美国的Phytophthora megasperma标准菌株为对照,将该7个菌株鉴定为Phytophthoramegasperma f.sp.glycinea。接种试验表明,该菌在不同大豆品种上致病力表现差异明显,而且对三叶草和紫花苜蓿不侵染。用MTT染色法定期检测埋在不同处理的土壤中的卵孢子活性。经一年后,所有处理中90%以上的卵孢子均处于活性状态;而温度对卵孢子的休眠期有较大的影响。     

Soybean root suberin and partial resistance to root rot caused by Phytophthora sojae

Phytophthora sojae is the causal agent of root and stem rot of soybean (Glycine max). Various cultivars with partial resistance to the pathogen have been developed to mitigate this damage. Herein, two contrasting genotypes, the cultivar Conrad (with strong partial resistance) and the line OX760-6 (with weak partial resistance), were compared regarding their amounts of preformed and induced suberin components, and to early events during the P. sojae infection process. To colonize the root, hyphae grew through the suberized middle lamellae between epidermal cells. This took 2 to 3 h longer in Conrad than in OX760-6, giving Conrad plants more time to establish their chemical defenses. Subsequent growth of hyphae through the endodermis was also delayed in Conrad. This cultivar had more preformed aliphatic suberin than the line OX760-6 and was induced to form more aliphatic suberin several days prior to that of OX760-6. However, the induced suberin was formed subsequent to the initial infection process. Eventually, the amount of induced suberin (measured 8 days postinoculation) was the same in both genotypes. Preformed root epidermal suberin provides a target for selection and development of new soybean cultivars with higher levels of expression of partial resistance to P. sojae.     

Real-time quantitative PCR assays for evaluation of soybean varieties for resistance to the stem and root rot pathogen Phytophthora sojae

Phytophthora root and stem rot caused by Phytophthora sojae is one of the most destructive disease of soybeans in the world. Effective management of the disease depends on selection and use of soybean varieties resistant to the disease. Fast and reliable procedures are vital to screen soybean varieties against the pathogen. Novel real-time quantitative (qPCR) assays were developed for both absolute and relative quantification of P. sojae in infected root tissues. QPCR assays were based on the detection of the internal transcribed spacer (ITS) gene of the pathogen and 18S ribosomal gene of the host plant. Absolute qPCR allowed the detection of as low as 10 femtograms (fg) of P. sojae DNA in soybean roots. Relative qPCR, employing the comparative threshold cycle (Ct) method, was effective and reliable for quantification of P. sojae DNA normalized to plant DNA in infected soybean root tissues. P. sojae DNA quantities detected in both qPCR assays had high correlations with disease severity index (DSI) ratings of soybean varieties. QPCR assays developed in this study were useful for determination of the levels of P. sojae DNA in different varieties of soybean and for evaluation of them for relative resistance to the pathogen.     

大豆和大豆疫霉菌共培养体系的构建

 本研究旨在建立一个适于分析大豆遭受大豆疫霉菌侵染后基因表达研究的互作体系。在比较了15个携带不同抗病基因的大豆基因型的组织培养情况和7个大豆疫霉菌分离物及其游动孢子单孢系的毒力变化的基础上,构建了大豆悬浮细胞和大豆疫霉菌游动孢子的共培养体系,进而分析了共培养过程中大豆细胞的活力和防御基因表达情况。结果表明,不同亲和力菌株的游动孢子引起的大豆细胞活力变化十分相似;非亲和菌株的游动孢子可诱导寄主防御基因的表达发生变化。此系统为深入开展大豆抗性机制研究提供了良好的平台。     

Phytophthora root rot of sweet pepper

Phytophthora capsici proved to be the causal agent of a root and crown rot of sweet pepper in the Netherlands.P. capsici was pathogenic on sweet pepper, tomato and sometimes on eggplant but not on tobacco Xanthi. Of these test plants only tomato was infected byP. nicotianae.No different symptoms in plants infected with eitherP. capsici orP. nicotianae were found. Dipping the roots of tomato and sweet pepper plants in a suspension ofP. capsici resulted in a more severe attack than pouring the suspension on the stem base.Resistance in tomato toP. nicotianae did not include resistance toP. capsici. A method to distinguishP. capsici fromP. nicotianae after isolation from soil is described. Both species were able to infect green fruits of tomato and sweet pepper.p. capsici survived in moist soil in the absence of a host for at least 15 months.Samenvatting Phytophthora capsici bleek de oorzaak te zijn van een voet-en wortelrot in paprika op twee bedrijven in 1977 in Nederland.P. capsici was pathogeen op paprika, tomaat en soms op aubergine maar niet op tabak Xanthi.P. nicotianae tastte van deze toetsplanten alleen tomaat aan. Verschillen in symptomen tussenP. nicotianae enP. capsici werden bij tomaat niet waargenomen.Het dompelen van de wortels in eenP. capsici suspensie gaf een ernstiger aantasting dan het begieten van de wortelhals met deze suspensie.Resistentie in tomaat tegenP. nicotianae bleek geen resistentie tegenP. capsici in te houden. P. capsici kan in grond worden aangetoond door groene paprikavruchten als vangsubstraat te gebruiken.P. capsici enP. nicotianae kunnen beide zowel vruchten van tomaat als paprika aantasten. P. capsici overleefde een periode van 15 maan den in vochtige grond waarop geen waardplant werd geteeld.     

大豆疫霉菌部分生物学特性及其药剂筛选研究

采用大豆疫霉菌在不同条件下菌丝生长速度法研究了大豆疫霉菌的部分生物学特性,并应用杀菌剂室内生测、盆栽药剂防效对药效作了评价。研究结果表明,大豆疫霉菌营养生长的最适温度为25～30℃;最适pH为6;光暗交替有利于该菌营养体的生长,在Rye或CA培养基上生长最快。室内药效测定结果表明,烯酰吗啉EC₅₀为0.165 4μg/mL,抑菌效果最好,甲霜灵、甲霜灵.锰锌和氟吗啉.锰锌的EC_{50分别为0.261 00、.451 0和0.984 2μg/mL,效果次之。盆栽试验结果表明,几种药剂在活体条件下对大豆疫病的防治效果较好,并有较长持效期。}     

大豆疫霉菌对大豆下胚轴侵染过程的细胞学研究

 接种后1.5~24h,用光镜和电镜研究了2个大豆品种与大豆疫霉菌Ps411的亲和性和非亲和性互作。观察结果表明,大豆疫霉菌对大豆下胚轴的侵染过程可分为侵入前、侵入、皮层组织中的扩展和进入维管束组织4个连续阶段。大豆下胚轴接种后在25℃保湿培养,1.5h后游动孢子即形成休止孢并萌发产生附着孢,3h后侵入表皮细胞,6h后进入皮层组织,24h后进入维管束组织。病原菌主要以侵染菌丝直接侵入表皮,表皮细胞间隙是主要侵入部位。皮层细胞是病原菌定殖和发展的主要场所,胞间菌丝侵入皮层细胞并形成吸器。在菌丝与寄主细胞接触部位的寄主细胞壁与质膜之间常有胞壁沉积物的形成。在抗病品种上病菌的侵染事件与感病品种基本一致,但不能形成正常的吸器,胞壁沉积物明显多于感病品种,菌丝在寄主组织内的扩展明显受到抑制。利用β-1,3-葡聚糖免疫金标记单克隆抗体进行的免疫细胞化学的研究表明,胞壁沉积物内含有大量的β-1,3-葡聚糖,在大豆疫霉菌菌丝壁中也存在β-1,3-葡聚糖。以上结果表明,病原菌的侵染可诱导抗病寄主细胞内β-1,3-葡聚糖迅速的合成与积累、并形成胞壁沉积物,以抵御病菌的侵染与扩展。     

Phytophthora root and crown rot of raspberry in Bulgaria

Root and crown rot of raspberry (Rubus idaeus L.) was observed in a plantation at the experimental station of small fruits in Kostinbrod, Bulgaria. Isolates ofPhytophthora spp. were obtained from diseased plants. Colony morphology, growth rates, features of asexual and sexual structures were studied and as a result twoPhytophthora species were identified:Phytophthora citricola Saw. andPhytophora citrophthora (R.E. Sm. & E.H. Sm.) Leonian. Their pathogenicity was confirmed in artificial inoculation experiments. The isozyme (-esterase) patterns ofP. citrophthora andP. citricola isolates from raspberry and from the collection of the CBS, Baarn the Netherlands were compared, using micro-gel electrophoresis. Both species are reported for the first time as pathogens of raspberry in Bulgaria. This is only the second report in phytopathological literature ofP. citrophthora on raspberry, the first being from Chile [Latorre and Munoz, 1993].     

Race distribution of Phytophthora sojae on soybean in Hyogo, Japan

Since 1987, Phytophthora root and stem rot of soybean [Glycine max (L.) Merr. cv. Tanbakuro], caused by Phytophthora sojae Kaufman and Gerdemann, has been increasing in the Sasayama, Nishiwaki, and Kasai regions in Hyogo, the most famous soybean (cv. Tanbakuro)-producing areas in Japan. In 2002 to 2004, 51 isolates (one from each field) of P. sojae were recovered from 51 fields in Hyogo. These isolates were tested for virulence on six Japanese differential soybean cultivars used for race determination in Japan, and three additional ones containing four Rps genes used in Indiana, USA. Race E was the most prevalent from 2002 to 2004, followed by races A, C, D, and four new races (proposed as races K, L, M, and N). Interestingly, none of the new races had high virulence on the Japanese differential cultivars, compared with other races in each area. One (race N) was avirulent on all six soybean differentials. There was a difference in race distribution on each of three individual areas; race E seemed to be a major component of the P. sojae population in Sasayama, whereas race A and the new race M were the most prevalent in Nishiwaki and Kasai, respectively. Rps6 (cv. Altona) and Rps1a + Rps7 (cv. Harosoy 63) were infected by 90.2% and 33.3% of all isolates, respectively. However, Rps1d (cv. PI103091) was not susceptible to any of the 51 isolates, nor was cv. Gedenshirazu-1. These two soybean cultivars were considered to be potential sources of resistance to breed new resistant cultivars with the desirable characteristics of cv. Tanbakuro for this region.     

江苏口岸截获大豆疫霉菌的鉴定

采用叶碟诱捕法从江苏口岸进境的大豆所携带的土壤中,共分离了疫霉12株,选取6个菌株,对病原菌进行了形态特征、致病性、寄主范围鉴定。结果表明,形态观察为疫霉属真菌,接种大豆后可出现典型的大豆疫病症状,且人工接种只侵染大豆、豇豆和菜豆等少数豆科植物。采用大豆疫霉的特异性引物检测表明,所有12个菌株均能扩增出分子量为330bp的特异性条带。结合形态和致病性测定,这些病原菌鉴定为大豆疫霉(Phytophthorasojae)。