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Molecular Detection of Phytophthora sojae
大豆疫霉菌ITS分子检测程序的建立及其应用

LIU Chun-Lai,YANG Ming-Xiu,WEN Jing-Zhi,
刘春来
,杨明秀,文景芝

微生物学通报 , 2007,
Abstract: An oligonucleotide primer pair was designed and synthesized after comparison and homological analysis of rDNA ITS sequences among Phytophthora sojae,its related Phytophthora species,and allied fungal and bacterial species from GenBank.PCR amplifications were carried out for 140 isolates including Phytophthora sojae.It showed that only isolates of Phytophthora sojae can be amplified and a special fragment of 288bp were produced by the primers.These primers were used to detect Phytophthora sojae in pure culture,inoculated diseased soybean plants,and inoculated soil samples.The detection protocol has good sensitivity to diseased tissues.
Races of Phytophthora sojae in Iran  [PDF]
Abbas Mohammadi,Azizollah Alizadeh,Mansore Mirabolfathey,Nasrin Nooras Mofrad
Pakistan Journal of Biological Sciences , 2008,
Abstract: Phytophthora root and stem rot of soybean is a destructive disease of soybean in Iran. Races 1 and 3 of pathogen have already been reported from two major growing regions of the crop, Lorestan and Golestan provinces. In a survey during 2004-2005, 142 isolates of P. sojae were recovered from infected plants and naturally infested soil samples using selective media and soybean leaf baiting technique. The majority of tested isolates (110 isolates) belonged to race one of P. sojae and 32 isolates belonged to race 3. ITS region of 23 isolates were amplified with specific primers Ps1 and Ps2. Sequences of this regions were similar to other gene banks sequences except two isolates from China. This survey showed low diversity in Iranian population of P. sojae.
豆秆黑潜蝇Melanagromyza sojae呋喃丹防治试验与危害损失率测定初报  [PDF]
杜金荣,洪礼铭
南京农业大学学报 , 1982, DOI: 10.7685/j.issn.1000-2030.1982.03.005
Abstract: 豆秆黑潜蝇是大豆的主要害虫之一。在江苏省一年发生5―6代,在夏大豆生长中后期的蛀茎率每年均达100%。经调查,江苏省目前尚不存在完全无蝇害的品种及完全无蝇害的地区。1981年作者承担了江苏省豆秆蝇协作组的豆秆黑潜蝇呋喃丹防治试验与危害损失率测定工作。研究结果表明全期防治蝇害的比不用药防治蝇害的豆株含虫量减少26.4―35.3%;大豆产量随豆株含虫量上升而减少,呈负相关,r=-0.882,达显著水平;因蝇害而引起的产量损失达20―30%。
豆秆黑潜蝇Melanagromyza sojae(Zehnter)田间分布型与抽样方法的初步研究  [PDF]
夏基康,王振荣,黎正宇
南京农业大学学报 , 1980, DOI: 10.7685/j.issn.1000-2030.1980.01.011
Abstract: 1979年7月20―22日、8月10―12日、9月10―12日分别从9、8、5块大豆田,应用分层随机抽样法进行豆秆黑潜蝇田间分布型调查。按豆田大小分?规划出各包括5至16个小区的区组。区组面积为20×100M至100×100M。每小区面积为25×25M(个别田的小区为20×20M)。在每小区内随机抽样4点,每点两排,每排5株。每区组抽样320株(8×40)至640株(16×40)(个别田为5×40株)。将豆株携回室内进行剥查。所获资料应用电子计算器(Casiofx―140)进行频次分布适合性测定。7月代的低密度虫口符合潘松分布,而8月、9月高密度虫口符合负二项分布与核心分布。样点含量大小以及豆株不同部位的虫口数影响样本的分布。一些扩散型指数(如Ca、I、L)以及平均拥挤度与平均数的回归关系在本文中应用并加以讨论。
大豆抗豆秆黑潜蝇(Melanagromyza sojae)性能的研究 Ⅰ.成虫生物学的观察  [PDF]
江苏省豆秆黑潜蝇研究协作组
南京农业大学学报 , 1981, DOI: 10.7685/j.issn.1000-2030.1981.03.006
Abstract: 豆秆黑潜蝇成虫在羽化后的第2天就有交尾,多数个体在羽化后3―4天内交尾,成虫产卵始迄期有5、6天,9天,少数个体长达24天,33天。产卵量在个体间的差异很大,最多的产卵396粒,雌成虫有卵巢小管16―26根,每小管的含卵粒数,最多达13粒.田间正在交尾雌虫的卵巢发育程度,个体间差异十分明显.成虫产卵对于不同生育期豆株有明显选择性.在V(?)植株和R_1、R_2植株上产卵为多.在野生大豆上产卵较少.成虫都产卵在豆叶背面距叶片基部(1厘米左右)的叶脉(主脉、支脉都可以)附近叶肉组织内.白天活动时喜欢趋集开白花的芝麻田。
Analysis of simple sequence repeats markers derived from Phytophthora sojae expressed sequence tags
Zhendong Zhu,Yunlong Huo,Xiaoming Wang,Junbin Huang,Xiaofei Wu
Chinese Science Bulletin , 2004, DOI: 10.1360/04wc0248
Abstract: Five thousand and eight hundred publicly available expressed sequence tags (ESTs) of Phytophthora sojae were electronically searched and 415 simple sequence repeats (SSRs) were identified in 369 ESTs. The average density of SSRs was one SSR per 8.9 kb of EST sequence screened. The most frequent repeats were trinucleotide repeats (50.1%) and the least frequent were tetranucleotide repeats (8.2%). Forty primer pairs were designed and tested on 5 strains of P. sojae. Thirty-three primer pairs had successful PCR amplifications. Of the 33 functional primer pairs, 28 primer pairs produced characteristic SSR bands of the expected size, and 15 primer pairs (45.5%) detected polymorphism among 5 tested strains of P. sojae. Based on the polymorphisms detected with 20 EST-SSR markers, the 5 tested strains of P. sojae were clustered into 3 groups. In this study, the SSR markers of P. sojae were developed for the first time. These markers could be useful for identification, genetic variation study, and molecular mapping of P. sojae and its relative species.
大豆抗豆秆黑潜蝇(Melanagromyza sojae Zehntner)性能的研究――Ⅱ.大豆产量损失和生理性状若干指标的测定  [PDF]
江苏省豆秆黑潜蝇研究协作组
南京农业大学学报 , 1984, DOI: 10.7685/j.issn.1000-2030.1984.03.004
Abstract: 本试验于1978―1982年分别在江苏省长江以北的4个地区进行。用辛硫磷、乐果、氧化乐果、呋喃丹等药剂控制植株体内虫量,经盆栽试验、小区试验、大区示范防治,各项结果一致表明,防治豆秆黑潜蝇为害可较大幅度地使大豆增产。防治区豆株的单株粒重、单株粒数、单株英数都显著地较未防治区豆株为高。在盆栽条件下,未经防治的豆株其叶片蒸腾强度、豆荚呼吸强度较经过防治的豆株为高。
Green fluorescent protein (GFP) as a vital marker for studying the interaction of Phytophthora sojae and soybean
XiaoRen Chen,BaoPing Cheng,XinLe Wang,SuoMeng Dong,YongLin Wang,XiaoBo Zheng,YuanChao Wang
Chinese Science Bulletin , 2009, DOI: 10.1007/s11434-009-0417-7
Abstract: Transgenic Phytophthora sojae strains that produce green fluorescent protein (GFP) were obtained after stable DNA integration using the Hsp70 promoter and the Ham34 terminator of Bremia lactucae. The expression of GFP during different developmental stages of P. sojae was observed using fluorescent microscopy. Based on this reporter system, the histopathologic events caused by the pathogen in soybean leaves, hypocotyls and roots were monitored. Meanwhile, the difference in resistance between different soybean cultivars against P. sojae was analyzed microscopically in roots. The results indicate that GFP can be stably expressed in zoosporangia, zoospores, cysts, hyphae and oospores of P. sojae. Using the GFP marker, the infecting pathogens in leaves, hypocotyls and roots of host could be distinctly visualized. The germ tube length of cysts germinating on the roots of resistant cultivar Nannong 8848 was longer than that on the roots of susceptible cultivar Hefeng 35. These results show for the first time that this eukaryotic reporter can be used in P. sojae as a stable and vital marker, allowing the study of genetics of this hemibiotrophic pathogen.
STUDIES ON RESISTANCE OF PHYTOPHTHORA SOJAE TO METALAXYL
大豆疫霉菌抗甲霜灵特性研究

Zuo YuHu,Hou JuMei,Kang ZheSheng,Chen ChangQing,Huang LiLi,
左豫虎
,侯巨梅,康振生,陈长卿,黄丽丽

菌物学报 , 2005,
Abstract: Phytophthora sojae developed the resistance to metalaxyl easily. The metalaxyl-resistant isolate (Mtr) of P. sojae was found in the wild-type isolates of P. sojae after treatment with metalaxyl. The resistance level of the metalaxyl-resistant isolate was over 870 times higher than that of the single zoospore progeny isolates of the wild-type parent isolate. The resistance to metalaxyl of Mtr isolate was inherited stably, and they did not show variation from the first zoospore progeny to the third zoospore progeny. The maintenance character of Mtr was not depended on metalaxyl. The resistance to metalaxyl of Mtr single zoospore isolate was not reduced and their zoospore progeny were not varied after 22-30 days growth on the metalaxyl free carrot agar medium (CA).
EMS mutagenesis analysis of Phytophthora sojae
大豆疫霉菌的EMS化学诱变

HA Xia,HU Zhong-Hui,WANG Lei,QUAN Jun-Li,SHAN Wei-Xing,
哈霞
,胡中慧,王蕾,权军利,单卫星

菌物学报 , 2010,
Abstract: The commonly used chemical mutagen EMS (ethylmethane sulfonate) was employed to create a mutant collection by treating the encysted zoospores of Phytophthora sojae and the spore germination was used to optimize the mutagenesis condition. A total of 640 single-oospore mutant lines were acquired, among which 50% of them showed a wide range of morphological changes. As to the production of oospores, 8.13% mutants produced more, 20.41% less, and 27.82% very few or no oospores as compared with the wild type, respectively, and 43.64% were similar to the wild type. The quality of the mutant collection was further confirmed by successful identification of 9 mutations in PsPMA1 (Plasma Membrane H+-ATPase 1) gene from 320 P. sojae mutants. Thus, the estimated maximum frequency of EMS induced mutation is 1 per 115kb of sequence. The created P. sojae mutant collection will be useful for functional genomic analysis of P. sojae.
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