黄淮地区大豆种质资源对疫霉根腐病的抗病性评价

【目的】建立一套大豆抗疫霉菌根腐病基因鉴别菌系,并用于分析大豆品种(系)是否含有抗病基因Rps1a、Rps1c和Rps1k。【方法】采用下胚轴接种法测定了125个大豆疫霉菌分离物的毒性组成,筛选了不同毒力公式的6个大豆疫霉菌并建立了该菌系,测定了黄淮地区55个主栽大豆品种(系)对该菌系的抗性并通过基因推导方法进行抗病基因分析。【结果】55个大豆品种(系)共产生18种反应型。抗病基因的推导结果表明,有2个品种可能含有Rps1c,没有鉴定到可能携带有Rps1a或Rps1k的大豆品种(系)。【结论】黄淮地区携带抗病基因Rps1a、Rps1c和Rps1k的大豆品种(系)并不多,且易感疫霉菌,因此需要及时进行抗性育种并积极推广。     

华南大豆种质对大豆疫霉根腐病的抗性分析

【目的】大豆疫霉根腐病是由大豆疫霉菌引起的严重影响大豆生产的世界性病害之一,选用抗病品种是控制该病最经济有效的措施。筛选抗大豆疫霉菌PGD1的优异抗源和多抗疫霉根腐病种质资源,推导大豆种质抗疫霉根腐病基因,为热带、亚热带地区大豆抗病育种提供有效抗源。【方法】采用下胚轴创伤接种法,利用在广东发现并分离的大豆疫霉菌PGD1菌株,接种鉴定主要来自广东、广西、福建、海南、湖南、江西和四川等华南省份631份大豆种质资源的抗病性,筛选抗大豆疫霉菌PGD1种质资源;再用其他6个不同毒力的大豆疫霉菌株接种鉴定抗大豆疫霉菌PGD1的种质,筛选多抗资源,通过基因推导方法分析抗病种质的抗病基因类型。【结果】631份大豆种质中有101份种质抗大豆疫霉菌PGD1,占鉴定种质的16.0%;73份为中间反应类型,占11.6%;457份表现感病,占72.4%。其中83份抗大豆疫霉菌PGD1的种质对其他6个不同毒力菌株Pm14、Pm28、PNJ1、PNJ3、PNJ4、P6497的侵染率分别为28.9%、34.9%、9.6%、66.3%、57.8%和10.8%。4份种质同时抗7个不同毒力的大豆疫霉菌株,分别为ZDD21538、ZDD21604、ZDD14286和明夏豆1号,占鉴定种质的4.8%。毒力频率为0的种质有15份,占鉴定种质的18.1%。83份大豆种质对7个不同毒力的大豆疫霉菌株共产生20种反应型,1种反应型与单个抗病基因的鉴别寄主Williams79反应型一致,可能含有抗病基因Rps1c;45份种质产生的9种反应型符合一些2个或2个以上已知抗病基因组合的反应型,这些种质可能含有已知抗病基因组合;38份种质共产生11种反应型既不同于任何含有单个已知抗病基因品种的反应型也不同于2个或2个以上已知抗病基因组合的反应型,它们可能含有新的抗病基因或基因组合。【结论】华南地区大豆种质中蕴藏着丰富的抗大豆疫霉菌PGD1和多抗大豆疫霉菌抗源,这些抗病种质可作为热带、亚热带地区大豆抗病育种的重要亲本和抗病基因定位的重要研究材料。     

黑龙江省东部大豆疫霉群体结构及其时空动态

为明确大豆疫霉群体结构及其时空变化规律,连续3年对黑龙江省东部4个具有代表性地块桦南县土龙山镇生产田Ht、桦南县曙光一队生产田Hs、黑龙江省农科院佳木斯分院大豆品种试验田Jn和农垦科学院大豆品种试验田Nk土样,采用叶碟诱捕法分离其中的大豆疫霉,利用一套含单个不同抗病基因的大豆近等基因系,通过国际通用下胚轴伤口接种法鉴定大豆疫霉生理小种。结果表明,379株大豆疫霉分别属于105种致病型,其中包括24个生理小种,1种IRT和80种新致病型,每种致病型优势度不明显。24个生理小种中,0、10、14和40号小种为中国首次报道。供试大豆疫霉群体对单个Rps基因的致病频率显示,在黑龙江省东部种植含有Rps7基因的大豆品种风险较高,而种植含有Rps1c、Rps1k和Rps3a基因的大豆品种较安全。随着时间推移,黑龙江省东部大豆疫霉单个菌株聚合对更多Rps基因毒性,单个菌株毒性增强。大豆疫霉群体毒性结构复杂,且随时间推移群体毒性也增强。空间上,生产田大豆疫霉群体较试验田群体毒性结构更复杂多样。     

一对单显性大豆抗疫霉根腐病基因的遗传分析及定位

[目的]大豆疫霉根腐病是大豆毁灭性的病害之一,防治该病最安全有效的方法是培育和利用抗性品种。迄今为止,已在大豆基因组上鉴定了21个大豆疫霉根腐病抗性基因,但是在中国只有少数基因(如Rps1c、Rps1k等)抗性有效。因此,急需发掘新的大豆疫霉根腐病抗性基因,以满足抗病育种的需要。‘大方六月早’对大豆疫霉菌的抗谱较广,是目前筛选出的优异抗源。[方法]采用下胚轴创伤接种方法进行抗病性鉴定,以品种‘大方六月早’为抗病亲本与品种‘矮脚早’配置杂交组合获得F2∶3代家系群体,并进行遗传分析,通过SSR(simple sequence repeat)标记对‘大方六月早’的抗性基因进行定位。[结果]该群体的抗性遗传分析表明:‘大方六月早’对大豆疫霉根腐病的抗性由1对主效基因控制,抗病对感病表现为显性,暂命名为RpsAH。使用SSR标记进行分子作图,抗病基因RpsAH被定位在大豆3号染色体(即N连锁群)上,距离标记Sat_166为4.1 cM。[结论]大豆品种‘大方六月早’对大豆疫霉菌株AH的抗性由1个单显性基因控制,该基因定位于大豆分子遗传图谱N连锁群上。     

新疆大豆疫霉菌毒力及大豆品种抗病性

对34株大豆疫霉菌(Phytophthora sojae)新疆分离物进行了毒力鉴定,发现新疆疫区大豆疫霉菌毒力结构复杂,不同地块之间、同一地块不同大豆疫霉菌分离物间都存在毒力差异.用7个不同毒力的分离物接种鉴定了19个新疆地区栽培大豆品种(系)的抗病性,其中10个品种(系)表现较好的抗性.     

中国大豆疫霉菌分布及毒力多样性研究

 通过大豆疫霉根腐病发生的田间调查、从病株和土壤中分离大豆疫霉菌 ,对大豆疫霉菌在我国的分布进行了研究 ,结果表明 ,除东北地区外 ,我国黄淮流域和长江流域也存在大豆疫霉菌。应用 13个鉴别寄主 ,对来自不同地区的 83个大豆疫霉菌分离物进行毒力鉴定 ,证明我国大豆疫霉菌存在丰富的毒力多样性。与植株分离物相比 ,土壤分离物的毒力多样性程度更高。对不同地区来源分离物的毒力组成比较表明 ,长江流域分离物的毒力多样性最丰富 ,其次是黄淮流域 ,而东北地区分离物的毒力组成相对简单。     

华南地区推广应用大豆品种对疫霉根腐病的抗性评价

【目的】评价华南地区大豆 Glycine max (L.)Merrill品种（系）及其亲本对7个大豆疫霉菌 Phytophthora sojae 的抗性.【方法】采用下胚轴创伤接种法鉴定67个品种（系）及其亲本对7个不同毒力的大豆疫霉菌菌株的抗性.【结果和结论】67个品种对7个不同毒力菌株PGD1、Pm14、Pm28、PNJ1、PNJ3、PNJ4、P6497的侵染率不同,接种大豆疫霉菌PGD1的侵染率最高,PGD1是华南地区发现并分离的大豆疫霉菌新种,抗PGD1的品种有华夏6号、粤夏2011-4、桂春6号、桂春10号、桂夏1号、浙春3号,占鉴定品种的9%;其次是Pm28和Pm14,接种PNJ1和PNJ4的侵染率较低;另外,对7个大豆疫霉菌菌株都表现感病的品种有29个,占鉴定品种的43.3%.华南地区抗或多抗大豆疫霉根腐病的大豆品种较少,需要加强大豆品种抗病种质资源筛选和抗病育种研究.     

南京大豆根腐病病原物的分离及毒性鉴定

对2005、2006年夏在南京农业大学江浦农场试验田发生的大豆根腐病,采用特异性PCR检测到发病组织中有大豆疫霉,经室内诱捕和分离,从发病田块的土壤和发病植株上共分离到4个大豆疫霉菌株PNJ1、PNJ2、PNJ3和PNJ4.用含有不同抗病基因的14个鉴别寄主测定这4个大豆疫霉菌株的毒力公式,PNJ1和PNJ2为1d,2,3b,3c,4,6,7;PNJ3为1a,1b,1c,1d,1k,2,3b,3c,5,7;PNJ4为1a,1b,1c,1d,1k,2,3b,3c,4,6,与国际上已经报道的大豆疫霉菌株的毒力公式不同,为新的生理小种.该研究可为抗病品种的选育及利用提供科学依据.     

与大豆疫霉根腐病抗性基因Rps1-c连锁的分子标记研究

以大豆疫霉根腐病近等基因系Williams和Williams79为材料,对它们的基因组DNA进行RAPD分析,160个10-nt随机引物中,只有引物OPQ-04在Williams79中扩增出特异条带,在Williams中未扩增出,且3次重复均获得一致结果。经进一步检测在其它抗病材料中也出现该特异条带,推测该特异条带与大豆抗疫霉根腐病抗性基因Rps1-c连锁,得到显性RAPD标记OPQ-04700。     

新疆大豆疫霉生物学特性及生理小种鉴定

从石河子、阿勒泰和伊犁大豆种植区采集大豆根腐病病样进行分离,共分离到5个疫霉菌株,经生物学特性测定,供试的新疆大豆疫霉各菌株在10～35℃均可生长,最适生长温度25℃左右;最适于菌丝生长的pH值为7左右;其菌丝在胡萝卜、V8汁培养基上生长较快,在V8汁、CA和大豆汁培养基上都能产生大量的卵孢子.生理小种鉴定表明,供试的大豆疫霉菌株分属4个不同的毒力公式,它们与当前国内外所报道的53个生理小种的毒力公式均有较明显差别,可能属于新的生理小种.     

Resistance Evaluation of Soybean Germplasm from Huanghuai Region to Phytophthora Root Rot

The aim of the study was to establish a set of differential strains and to identify soybean resistant genes to Phytophthora root rot and then to apply those strains for analysis of the resistant genes Rps1a, Rps1c, and Rps1k that soybean cultivars or lines may carry. Virulence formula of 125 Phytophthora sojae isolates were determined using the hypocotyls inoculation technique, the strains, which includ 6 isolates with different virulence formulas, were applied to identify the resistance of 55 soybean cultivars or lines and resistant genes were analyzed using the gene postulating procedure. Eighteen reaction types occurred in 55 cultivars or lines and results of gene postulation indicated that 2 cultivars or lines probably carried gene Rpslc and no cultivar may carry genes Rpsla or Rpslk. A few of soybean cultivars or lines from Huanghuai Region carry Rps genes Rpsla, Rpslc and Rpslk and tend to infect by P. sojae, so resistant cultivars or lines need to be bred and popularized actively.     

新疆大豆疫霉的来源分析

应用SSR标记对新疆地区大豆疫霉菌（Phytophthora sojae Kaufmann ＆ Gerdemann）的遗传多样性和与美国、黑龙江、福建等3个不同地理来源大豆疫霉菌的遗传关系进行研究。结果表明,新疆地区大豆疫霉菌遗传多样性相对较低,遗传背景单一,推断为外来生物。与大豆疫霉菌美国分离物和福建分离物相比,新疆分离物与黑龙江分离物的遗传关系最近,表明新疆大豆疫霉菌可能从黑龙江传人。     

Identification, Genetic Analysis and Mapping of Resistance to Phytophthora sojae of Pm28 in Soybean

Phytophthora sojae Kanfman and Gerdemann (P.sojae) is one of the most prevalent pathogens and causes Phytophthora root rot,which limits soybean production worldwide.Development of resistant cultivars is a cost-effective approach to controlling this disease.In this study,127 soybean germplasm were evaluated for their responses to Phytophthora sojae strain Pm28 using the hypocotyl inoculation technique,and 49 were found resistant to the strain.The hypocotyl of P1,P2,F1,and F2:3 of two crosses of Ludou 4 (resistant)×Youchu 4 (susceptible) and Cangdou 5 (resistant)×Williams (susceptible) were inoculated with Pm28,and were used to analyze the inheritance of resistance.The population derived from the cross of Ludou 4×Youchu 4 was used to map the resistance gene (designated as Rps9) to a linkage group.932 pairs of SSR primers were used to detect polymorphism,and seven SSR markers were mapped near the resistance gene.The results showed that the resistance to Pm28 in Ludou 4 and Cangdou 5 was controlled by a single dominant gene Rps9,which was located on the molecular linkage group N between the SSR markers Satt631 (7.5 cM) and Sat_186 (4.3 cM).     

Impact of Crop Rotation on Pathotype and Genetic Structure of Phythophthora sojae in Fields

To estimate the impact of crop rotation on the pathotype and genetic structure ofPhythophthora sojae in fields, 372 isolates ofP. sojae were obtained from long-term localisation experimental fields in Heilongjiang Province of China. The hypocotyl inoculation method was used to characterize the virulence ofP. sojaeon 13 differential cultivars, and the amplified fragment length polymorphism (AFLP) technique was used to analyze difference in the genetic structure ofP. sojae. The results indicated that an abundant diversity of genetic structures and pathotypes ofP. sojae, a more uniform distribution of pathotypes and less dominance of pathotypes occurred in corn-soybean and wheat-soybean rotation fields than in a continuous soybean mono-cropping field. These findings suggested thatP. sojae did not easily become the dominant race in rotation fields, which maintain disease resistance in soybean varieties. Therefore, the results of this study suggested that Phytophthora stem and root rot of soybeans could be effectively controlled by rotating soybeans with non-host crops of corn and wheat.     

大豆疫霉基因组SSR标记开发

 【目的】开发大豆疫霉基因组SSR标记,为从分子水平深入研究大豆疫霉及其近缘种提供一种理想的分子标记。【方法】用FPCR软件从大豆疫霉全基因组序列中查询SSRs,选择合适的SSR序列用Primer5.0软件设计引物。【结果】从发现的1 234个含有2～4个碱基重复单元的完全SSRs中选出260段设计引物,经10个大豆疫霉分离物基因组DNA检测,有213对(81.9%)扩增出SSR特征条带,其中114(53.5%)对引物扩增出多态性。通用性检测表明,14.6%～28.6%引物分别在选择的8个疫霉种中有效扩增。基于10个SSR标记数据进行聚类分析,结果表明表明这些标记可以完全区分大豆疫霉及其它疫霉。【结论】大豆疫霉基因组SSR标记具有高多态性,是大豆疫霉遗传变异、遗传多样性、及遗传图谱构建等研究理想工具。部分大豆疫霉基因组SSR标记在其近缘种中具有通用性,可以用于疫霉菌的系统进化、鉴定、区分及开发近源种的SSR标记。特别开发的SSR标记,在大豆疫霉基因组中具有准确的位置,将极大地方便大豆疫霉菌功能基因的定位和克隆,以及进行疫霉菌的比较基因组的研究。     

Evaluation of Soybean Germplasm from Provinces in Northeast China for Resistance to Phytophthora sojae

Soybean Phytophthora root rot (Phytophthora sojae) is a severe disease all over the world. Soybean germplasm from central and southern China for resistance has been evaluated by American researchers on a large scale. P. sojae has been found frequently in northeast of China in recent years, but not systematic evaluation of soybean germplasm for resistance has occurred there. By means of hypocotyl inoculation, 922 cultivars/lines from northeast of China were screened and evaluated for their response to race 1, and 25 of P.sojae. Generally resistance was less frequent in northeast of China than in central and southern China. Five cultivars/lines were identified that confer resistant responses to race 1,3,8,25 and four additional isolates of P.sojae. These cultivars/lines may provide valuable sources of resistance for future breeding programs.     

大豆疫霉(Phytophthora sojae)单卵孢株毒性纯系的建立

将培养30d的大豆疫霉1号生理小种卵孢子在适宜条件下诱导萌发,随机挑取30个单卵孢株进行纯培养和毒力测定,在测定结果与亲代一致(1号生理小种)的菌株中随机选取1个单卵孢株重复上述试验。试验经过连续两代单卵孢株分离培养,获得了毒力稳定表达的大豆疫霉1号生理小种单卵孢株毒力纯系,为开展大豆疫霉毒性遗传和变异的分子生物学研究提供了研究材料。