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大豆疫霉Phytophthora sojae卵孢子在黑龙江省土壤中的越冬存活率
Overwintering survival rate of Phytophthora sojae oospores in soils in Heilongjiang Province
 [PDF]

陈秋明,肖彩霞,孙欠欠,文景芝,Chen Qiuming,Xiao Caixia,Sun Qianqian,Wen Jingzhi
- , 2015, DOI: 10.13802/j.cnki.zwbhxb.2015.01.011
Abstract: 为探究大豆疫霉 Phytophthora sojae卵孢子在黑龙江省土壤中的越冬存活率及其与所处土壤深度和媒介的相关性,以增强型绿色荧光蛋白为报告基因,将培养基及病残体中的大豆疫霉卵孢子分别接种到试验田框栽土壤表层下不同深处,检测其卵孢子的越冬存活率,同时在框栽中定量播种不含任何已知抗疫霉根腐病基因的大豆品种Sloan(rps),苗期调查其发病率。结果表明,大豆疫霉卵孢子在黑龙江省土壤中的适生性较强,可在5~15 cm深度土壤中安全越冬,越冬存活率高达81.67%~96.33%。卵孢子越冬存活率与其所处的越冬媒介关系不大,而与土壤深度有关。在5~15 cm范围内,随着土壤深度的增加,卵孢子越冬存活率增加。处于深层土壤中的卵孢子更容易打破休眠,进入萌发前的萌动状态。各处理间卵孢子越冬存活率的显著性差异并未在发病率上表现出来,说明除了土壤深度外,还有其它因素影响发病率。
In order to investigate the associations between the survival rate of overwintering oospores of Phytophthora sojae and the soil depth and media in which the oospores were in Heilongjiang Province. The oospores in the media and diseased soybean tissues were inoculated at different depths below the soil surface in experimental fields under natural conditions. Some samples were collected randomly to measure the survival rate of oospores under a fluorescent microscope. Meanwhile, the seeds of susceptible soybean cultivar Sloan containing no R gene (rps) to Phytophthora root and stem rot were planted quantificationally in the frames to measure the incidence at seedling stage. The results showed that the oospores of P. sojae could overwinter with a survival rate of 81.67%-96.33%, indicating that they were adapted to the soil environment in Heilongjiang Province. The survival rate of overwintering oospores was related to the soil depth where oospores located, and had little relationship with the media. With the depth increasing, the survival rate of overwintering oospores increased. Oospores that located in deep soils were more likely to break dormancy to germinate. The significant differences in the survival rate of overwintering oospores between different treatments did not express in the seedling incidence, indicating that, in addition to the soil depth, there were other factors influencing the incidence of this disease.
Genetic relationships between Anhui and Heilongjiang populations of Phytophthora sojae assessed by SSR markers
安徽和黑龙江省大豆疫霉群体遗传结构的SSR分析

WANG Zi-Ying,WANG Zhao-Xia,SHEN Jie,LU Hong-Xia,
王子迎
,王朝霞,沈洁,鲁红侠

菌物学报 , 2009,
Abstract: The genetic diversity of two geographic populations of Phytophthora sojae Kauf. & Gerd. from Anhui and Heilongjiang Provinces was determined using the molecular marker of simple sequence repeats (SSRs). Genetic variation was analyzed for 83 isolates of P. sojae. By using 20 pairs of SSR primers, a total of 109 polymophic bands (alleles) were amplified at an average of 5.5 bands per pair of primers. Genetic similarity analysis showed that there were obvious differences between Helongjiang and Anhui populations. Cluster analysis with an unweighted pair group method revealed that the 83 isolates of P. sojae were clearly separated into seven clustering groups (genotypes) at a level of 80% similarity. We also found that isolates from Heilongjiang had lower genotypic diversity than those of Anhui. In addition, three particular P. sojae genotypes were only found in Anhui population and two particular genotypes were only found in Heilongjiang population. In conclusion, the results in this study do not support the hypothesis that P. sojae in Anhui has immigrated from Heilongjiang.
黑龙江省东部大豆疫霉群体遗传多样性时空动态 Spatial-temporal dynamics of genetic diversity of Phytophthora sojae in eastern Heilongjiang Province  [PDF]
文景芝,张斌,田苗,揭岩,赵钰琦,高新颖,宫丽娟,顾鑫,丁俊杰,陈宇飞
- , 2018,
Abstract: 为明确大豆疫霉群体遗传多样性时空变化规律,采用简单重复序列(Simple sequence repeat,SSR)标记技术,分析2014~2017年黑龙江省东部生产田和试验田大豆疫霉群体遗传多样性。选用8对SSR引物扩增547株供试大豆疫霉,获得123个条带,其中110个为多态性条带,占89.43%,基因多态性位点主要集中在700、450和350 bp,在450 bp处基因多样性复杂;遗传变异分析表明,随时间推移,黑龙江省东部大豆疫霉群体内基因变化极显著,遗传多样性复杂,基因变异频率升高,群体明显进化。试验田大豆疫霉群体遗传多样性丰富,基因变异频率略高于生产田。
Avirulent genes of Phytophthora sojae were detected by rapid molecular method
大豆疫霉菌无毒基因的快速检测

FAN Li,
范黎

微生物学通报 , 2012,
Abstract: 大豆疫霉菌Phytophthora sojae可引起大豆疫霉根腐病,是影响大豆生产的毁灭性病害。全球每年由于大豆疫霉根腐病导致的直接经济损失高达十几亿美元,该菌流行于我国东北大豆产区和福建等地并引起严重病害1]。
Identification of a repetitive sequence element for DNA Fingerprinting in Phytophthora sojae
大豆疫霉菌一个DNA指纹分析重复序列探针的鉴定

Lihua Yin,Qinhu Wang,Feng Ning,Xiaoying Zhu,Yuhu Zuo,Weixing Shan,
殷丽华
,王秦虎,宁峰,朱晓莹,左豫虎,单卫星

微生物学报 , 2010,
Abstract: Abstract: Objective] Establishment of DNA fingerprinting in Phytophthora sojae and an analysis of genetic relationship of Heilongjiang and Xinjiang populations. Methods] Bioinformatics tools were used to search repetitive sequences in P. sojae and Southern blot analysis was employed for DNA fingerprinting analysis of P. sojae populations from Heilongjiang and Xinjiang using the identified repetitive sequence. Results] A moderately repetitive sequence was identified and designated as PS1227. Southern blot analysis indicated 34 distinct bands ranging in size from 1.5 kb-23 kb, of which 21 were polymorphic among 49 isolates examined. Analysis of single-zoospore progenies showed that the PS1227 fingerprint pattern was mitotically stable. DNA fingerprinting showed that the P. sojae isolates HP4002, SY6 and GJ0105 of Heilongjiang are genetically identical to DW303, 71228 and 71222 of Xinjiang, respectively. Conclusion] A moderately repetitive sequence designated PS1227 which will be useful for epidemiology and population biology studies of P. sojae was obtained, and a PS1227-based DNA fingerprinting analysis provided molecular evidence that P. sojae in Xinjiang was likely introduced from Heilongjiang.
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.
土壤环境对大豆疫霉Phytophthora sojae卵孢子萌动的影响
Effects of soil environment on activation of oospores of Phytophthora sojae
 [PDF]

所冰,崔人方,田苗,文景芝,Suo Bing,Cui Renfang,Tian Miao,Wen Jingzhi
- , 2015, DOI: 10.13802/j.cnki.zwbhxb.2015.03.013
Abstract: 为明确土壤环境对大豆疫霉Phytophthora sojae卵孢子萌动的影响,将增强型绿色荧光蛋白标记的大豆疫霉卵孢子以2 500个卵孢子/g干土的比例接种于菌黑土中,荧光显微镜下计数卵孢子的萌动率以明确其最适宜的土壤温度和含水量;在此基础上,从5种类型土壤和5种轮作体系土壤中筛选适宜卵孢子萌动的土壤环境.结果表明,25 ℃土壤温度和30%土壤含水量最适合大豆疫霉卵孢子萌动,萌动率为95.78%;黑土和盐碱土分别是最适合和最不适合卵孢子萌动的土壤类型,萌动率分别为94.94%和14.67%;卵孢子萌动率与土壤有机质含量、pH 值和Ca2+含量间明显相关性.大豆连作田和玉米连作田土壤适合卵孢子萌动,萌动率为96.33%和95.00%,小麦连作田土壤不适合卵孢子萌动,萌动率仅为39.33%.
In the present study, the oospores marked by the enhanced green fluorescent protein (EGFP) were inoculated at a certain proportion to the Mollisols through a sterile Eppendorf tube to determine the most suitable soil temperature and water content by calculating the activation rate of oospores under the fluorescent microscope. The same method was used to measure the oospore activation rate in five types of soils and five types of crop rotation soils under the most suitable soil temperature and soil water content to determine the most suitable soil environment. The results showed that the soil temperature of 25 ℃ and the soil water content of 30% were the most suitable for oospore activation, with an activation rate of 95.78%. Mollisols and Halosols were the most suitable and unsuitable soil types, respectively, for the activation of oospores, with an activation rate of 94.94% and 14.67%, respectively. There was no correlation between the activation rate of oospores and pH value, organic matter and Ca2+ content of the soil. Also, the soil from soybean mono-cropping field and corn mono-cropping field were more suitable for the activation of oospores than that from wheat mono-cropping field, with an activation rate of 96.33%, 95.00% and 39.33%, respectively.
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.
Genetic relationships among Chinese and American isolates of Phytophthora sojae by ISSR markers
中国和美国大豆疫霉群体遗传结构的ISSR分析

Ziying Wang,Yuanchao Wang,Zhengguang Zhang,Xiaobo Zheng,
王子迎
,王源超,张正光,郑小波

生物多样性 , 2007,
Abstract: The genetic diversity of three geographic populations of Phytophthora sojae from China and the United States was determined using technique of inter-simple sequence repeat(ISSR).A total of 102 repro-ducible ISSR fragments were scored among 111 individuals,of which 88(86%) were polymorphic.Analysis of genetic variation showed that there existed higher genetic variation in the United States population in comparison to the Chinese populations.Nei's genetic identity and principal component analysis indicated that the populations of Fujian and the United States are closer to each other than to Heilongjiang populations.Shannon-Wiener diversity index revealed that the United States populations have a higher genetic diversity than that of the Chinese populations.These data support the hypothesis that P.sojae in the United States might not have been introduced from China.
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