普通小麦-柔软滨麦草易位系M97抗条锈基因的遗传分析和分子作图

为了明确M97抗条锈性遗传规律,在苗期用7个小麦条锈菌系对M97与感病品种铭贤169的杂交后代F1、F2、F3和BC1代进行抗条锈性遗传分析,并对M97抗Sun11-4的抗条锈基因进行SSR分子标记。M97对Sun11-4和Sun11-11的抗病性均由1对显性基因控制,对CY29、CY30、CY33的抗病性由1显1隐2对基因共同控制,对CY31的抗病性由2对显性基因独立或重叠作用控制。以接种Sun11-4的F2代分离群体构建作图群体,筛选到Xwmc222、Xwmc147、Xbarc229和Xwmc339等4个与抗病基因连锁的SSR标记,其遗传距离分别为3.4、4.8、7.6和12.1 cM。将该抗病基因定位于小麦1DS染色体,且该基因不同于已知的抗条锈基因,暂命名为YrM97。用YrM97两侧遗传距离最近的2个标记Xwmc222和Xwmc147对42个黄淮麦区主栽小麦品种进行分子检测,仅有9.5%的品种具有与YrM97相同的标记位点。     

黄淮麦区小麦全蚀病菌群体的遗传多样性分析

为了解小麦全蚀病病菌的群体组成和遗传多样性,采用8对多态性较好的微卫星标记,对我国黄淮麦区的116个小麦全蚀病菌株进行了分析。8个SSR标记的平均等位基因数为4.25个,多态性信息含量的平均值为0.66。供试菌株的平均有效等位基因数和基因遗传多样性指数分别为1.46和0.27,漯河群体的遗传多样性水平最高,周口群体最低。不同群体间遗传距离均较小,为0.0199~0.1153,其中徐州和周口群体间的遗传距离相对较大,而周口和驻马店群体间的遗传距离相对较小。分子方差分析结果显示,小麦全蚀病菌群体间和群体内均存在着一定的遗传分化,群体间的遗传变异占总变异的10%,群体内遗传变异占90%。6个群体间的基因流为3.5。根据SSR多态性,对来源不同菌株的UPGMA聚类分析结果显示,小麦全蚀病菌群体结构与地理来源有一定的关系。     

水稻稻瘟病抗性QTL的定位分析

 稻瘟病是危害水稻最严重的病害之一。以抗稻瘟病的云南省地方品种魔王谷（MWG）和感稻瘟病的湖北省审定品种鄂晚8号（EW8）为亲本材料,构建双单倍体分离群体（DH）。利用22个菌株对亲本材料MWG/EW8进行致病性鉴定,从中筛选到5个毒性不同的鉴别菌株用于考察DH群体的稻瘟病抗性,构建包含120对SSR标记的分子遗传连锁图进行数量性状位点（QTL）的分析,鉴定出3个抗性QTL,均位于第6染色体长臂RM541附近, 3个QTL对抗病表型的贡献率介于7.7%~15.2 %之间,3个QTL的抗病等位基因均源自亲本MWG。     

小麦农家种红蚰麦抗白粉病遗传分析及SSR分子标记

 为明确小麦农家种红蚰麦抗白粉病的遗传基础,对红蚰麦和豫麦13的杂交F₂代群体进行了遗传分析,结果表明红蚰麦携带1对显性的抗白粉病基因(暂命名为Pmhym)。利用SSR标记和F₂代分离群体分组分析法,将该基因定位在7B染色体的长臂上,与3个微卫星标记Xwmc232、Xgwm577和Xwmc526连锁,遗传距离分别是14.3、25.6和57.2cM。分子标记分析表明该基因不同于已有被定位在7BL上的Pm5系列复等位基因,因而推测Pmhym是1个新的抗白粉病基因。上述结果将为开展Pmhym基因的精细定位奠定基础。     

基于KASP技术的小麦条锈菌SNP分子标记开发与评价

为开发用于小麦条锈菌Puccinia striiformis f. sp. tritici群体遗传研究的竞争性等位基因特异性PCR-单核苷酸多态性(kompetitive allele specific PCR-single nucleotide polymorphism,KASPSNP)标记,以中国小麦条锈菌流行小种CYR32的基因组为参考,与美国小麦条锈菌流行小种PST78和印度小麦条锈菌流行小种38S102的基因组进行比对,根据比对到的SNP位点设计KASP-SNP引物,用64个中国小麦条锈菌标样对其进行筛选,同时用13对多态性引物组成的简单重复序列(sim‐ple sequence repeat,SSR)分子标记分析这64个标样,并利用Powermarker 3.25和Structure 2.3软件通过多态性指数和群体遗传结构分析来评价KASP-SNP和SSR两种分子标记。结果显示,共比对到29 929个SNP位点,设计出462对KASP-SNP引物,经64个中国小麦条锈菌标样筛选到43对多态性较好的引物,所开发的这43对KASP-SNP引物多态性信息含量指数平均为0.346,基因多样性指数平均为0.420,而SSR引物的2种指数分别为0.237和0.265,前者较后者分别高出46.0%和58.5%。2种标记结果的群体遗传结构分析可得到类似结果,最佳聚类数K值都为4,云南菌系是遗传结构相对最简单的菌系,湖北菌系是遗传结构相对最复杂的菌系,但个别菌株的遗传划分存在较大差异。表明本研究开发的KASP-SNP分子标记多态性较SSR分子标记更加丰富,具有较好的应用前景。     

小麦山农4143抗黑胚病遗传分析及抗性遗传位点检测

黑胚病是小麦生产的重要籽粒病害,麦根腐平脐蠕孢(Bipolaris sorokiniana)是黑胚病的主要致病菌.为分析小麦抗黑胚病遗传规律并检测抗性位点,本研究以抗黑胚病小麦品系山农4143与感病品系宛原白1号的F7代重组自交系(RIL)群体为材料,于2018～2019年在3个试验点种植,采用“孢子液喷洒、套袋保湿”...     

源于华山新麦草抗条锈病基因 YrH122遗传分析和SSR标记

 H122是1个通过杂交和回交选育的普通小麦-华山新麦草易位系。为明确其抗条锈病基因及遗传特点,建立抗病基因SSR标记,利用我国小麦条锈菌流行小种CYR29、CYR31、CYR32、CYR33和致病类型Su11-4、Su11-11对H122进行苗期抗性鉴定,根据鉴定结果选用CYR32、CYR33和Su11-4对其与铭贤169杂交F1、F2及BC1代进行了遗传分析,同时应用258对SSR引物对将H122/铭贤169 F2代接种Su11-4的185个单株构建的作图群体进行了PCR扩增和电泳分析。结果表明,H122对供试小种均表现免疫或近免疫,对CYR32的抗病性由1对显性基因控制,对CYR33的抗病性由1对隐性基因控制,对Su11-4的抗病性亦由1对显性基因控制,将其暂命名为YrH122。筛选到3个与YrH122连锁的SSR标记Xbarc229、Xwmc339和Xwmc93,遗传距离分别为7.7、4.3和11.0 cM,并将该基因定位于小麦染色体1DL上。SSR标记回检显示,YrH122来源于华山新麦草。通过基因来源、分子检测及染色体位点比较,YrH122可能是1个不同于目前已知抗条锈病基因的新基因。     

源于柔软滨麦草抗小麦条锈病基因YrElm的遗传分析与SSR标记

 M852-1是由柔软滨麦草和普通小麦7182经杂交和回交培育的易位系。苗期抗病性鉴定结果表明,M852-1对CYR29、CYR31、CYR32、CYR33、Su11-4、Su11-7和V26等7个中国小麦条锈菌主要生理小种或新的致病类型均表现免疫至高抗,是一个较好的抗条锈资源材料。用条锈菌流行小种CYR33对M852-1与铭贤169杂交F₁、F₂、F₃和BC₁代进行抗性鉴定与遗传分析,发现M852-1对CYR33的抗条锈性由1对隐性基因控制,暂定名为YrElm。以F₂代分离群体构建作图群体,利用集群分离分析法,筛选到与YrElm连锁的5个SSR标记:Xcfd35、Xgwm161、Xwmc630、Xgwm533和Xcfd34,并将YrElm定位于小麦染色体3DS上。YrElm两侧最近2个SSR标记Xcfd35与Xgwm161的遗传距离分别为6.5 cM和4.2 cM。抗锈性鉴定、系谱分析以及分子标记检测结果表明,该抗病基因来源于柔软滨麦草。综合基因来源、分子检测及染色体位点等方面的分析,认为YrElm可能是一个新的抗条锈病基因。用该基因两侧最近两个标记Xcfd35和Xgwm161 检测68个甘肃和黄淮麦区小麦品种(系),10个(14.7%)品种能扩增出与M852-1相同的条带。进一步进行抗病性及系谱分析表明,这10个品种均不含YrElm。本研究结果为利用YrElm进行分子标记辅助育种和进一步的精细定位奠定了基础。     

烟草疫霉SSR分子标记的开发与初步应用

 为开发烟草疫霉的SSR分子标记,利用MISA软件搜索烟草疫霉基因组序列中的 SSR位点,共发现1 311个SSR位点,优势SSR位点为二核苷酸和三核苷酸,分别占总 SSR 位点的56.45%和39.36%;根据分析到的SSR位点使用Primer 5.0软件设计48对SSR引物,以7株烟草疫霉的DNA 为模板对这些引物进行筛选,共获得扩增条带清晰且具有多态性的SSR引物20对,然后使用其中的6对SSR对32株烟草疫霉进行UPGMA聚类分析,遗传相似系数在 0.60~1.00 之间,在相似系数0.70水平上,可将其划分为3个类群,类群I包含29个菌株,类群II包含1个菌株,类群III包含2个菌株,显示出较低的遗传分化水平。这些SSR引物的开发将为研究我国烟草疫霉的遗传多样性、群体遗传结构以及遗传图谱构建等奠定基础。     

普通小麦-柔软滨麦草易位系M852-1抗条锈基因的遗传分析和分子作图

 M852-1是经杂交和回交培育的普通小麦-柔软滨麦草易位系,苗期对我国小麦条锈菌流行小种均表现良好抗性。为明确其抗条锈性遗传规律,本研究选用条锈菌流行小种（类型）CYR29、CYR32、CYR33和Su11-7的单孢菌系对其与铭贤169杂交F₁、F₂、F₃及BC₁代群体进行遗传分析, 同时应用420对SSR引物对接种CYR32的M852-1/铭贤169 F₂代144个单株作图群体进行抗病基因定位。结果表明,M852-1对供试小种均表现免疫或近免疫,对CYR29的抗锈性由1对显性基因控制,对CYR32、CYR33和Su11-7的抗锈性均由1对隐性基因控制。筛选到3个与抗CYR32基因连锁的SSR标记Xbarc124、Xbarc200和Xgwm429,遗传距离分别为6.3、5.6 和 9.7 cM。根据SSR标记锚定性将该基因定位于小麦2BS染色体,暂命名为YrM852。基因来源、分子标记检测及染色体位点分析表明,YrM852很可能是1个不同于目前已知抗条锈病基因的新基因。     

Identification of quantitative Loci for tolerance to barley yellow dwarf virus in oat

ABSTRACT Molecular markers linked to quantitative trait loci conditioning tolerance to barley yellow dwarf virus (BYDV) were identified in oat (Avena sativa) using amplified fragment length polymorphism (AFLP) analysis. Near-isogenic and recombinant inbred lines (NILs and RILs, respectively) derived from a cross of Clintland64 (BYDV-sensitive) and IL86-5698 (BYDV-tolerant) were evaluated for their responses to an Illinois isolate of the PAV strain of BYDV. Individual markers identified in the analysis of the NILs explained up to 35% of the variability seen in the tolerance response. Single-point analysis of the marker data from the RIL population identified 24 markers in three linkage groups that were associated with tolerance to BYDV infection at P /= 3.0. These loci explained about 50% total of the variation in BYDV tolerance in multimarker regression analysis in both years. The BYDV tolerance loci A, C, E, and R were mapped to hexaploid oat restriction fragment length polymorphism linkage groups 2, 8, 36, and 5, respectively, by analyzing the segregation of the AFLP markers in the Kanota x Ogle RIL population.     

以Taichung29为背景的小麦抗条锈病近等基因系转育进展

自1988年起,系统开展了以Taichung29为背景的小麦抗条锈病近等基因系转育及其基础性研究。采用回交法和系谱法相结合的转育方法,以春性品种Taichung29为轮回亲本作母本,分别与25个抗性供体即中国小麦条锈病菌鉴别寄主和国际上重要的抗条锈基因载体品种杂交、回交和自交。通过基因推导分析、单体分析和SSR标记技术检测目的基因,选育抗条锈近等基因系,现已获得重要进展,成功选育出8个以Taichung29为背景的抗条锈病单基因近等基因系,即Taichung29*6／Yr1、Taichung29*6／Yr2、Taichung29*6／Yr5、Taichung29*6／Yr7、Taichung29*6/Yr9、Taichung29*6／Yr10、Taichung29*6/YrSpP、Taichung29*6/YrKy2。另有9个组合转育获得343个抗性稳定株系,正检测其目的基因,3个组合转至BC₆F₃,自交纯合筛选抗性稳定株系,5个组合转至BC₅,继续回交转育。     

Development of Co-Dominant Amplified Polymorphic Sequence Markers in Rice that Flank the Magnaporthe grisea Resistance Gene Pi7(t) in Recombinant Inbred Line 29

ABSTRACT Pi7(t), a dominant blast resistance gene derived from the rice cultivar Moroberekan, confers complete resistance against the fungal pathogen Magnaporthe grisea. Pi7(t) previously was positioned on chromosome 11 by restriction fragment length polymorphism (RFLP) mapping of a recombinant inbred line population. One derivative of this population, recombinant inbred line (RIL)29, was designated as the representative line for Pi7(t). A segregating F2 population was created from RIL29 in order to determine the location of Pi7(t). The new mapping data indicate a position for Pi7(t) 30 centimorgans distal to the original location. Pi7(t) shares a common position with the previously mapped Pi1 M. grisea resistance gene. RIL29 carries DNA not derived from either parent used to create the RIL population at the newly assigned Pi7(t) locus. RFLP analysis has identified a possible donor source.     

Mechanism and molecular markers associated with rust resistance in a chickpea interspecific cross (Cicer arietinum × Cicer reticulatum)

A gene that controls resistance to chickpea rust (Uromyces ciceris-arietini) has been identified in a recombinant inbred line (RIL) population derived from an interspecific cross between Cicer arietinum (ILC72) × Cicer reticulatum (Cr5-10), susceptible and resistant to rust, respectively. Both parental lines and all RILs displayed a compatible interaction but differed in the level of infection measured as Disease Severity (DS) and Area Under the Disease Progress Curve (AUDPC). Histological studies of the seedlings of resistant parental Cr5-10 line revealed a reduction in spore germination, appressorium formation, number of haustoria per colony and colony size, with little host cell necrosis, fitting the definition of partial resistance. A Quantitative Trait Locus (QTL) explaining 31% of the total phenotypic variation for DS in seedlings and 81% of the AUDPC in adult plants in the field was located on linkage group 7 of the chickpea genetic map. The AUDPC displayed a bimodal distribution with high frequency of susceptible lines and both the AUDPC and markers showed the same distorted segregation. Consequently, it was hypothesised that a single dominant gene (proposed as Uca1/uca1) controlled resistance to rust in adult plants. This allowed us to locate the gene on the genetic linkage map. Two Sequence Tagged Microsatellite Sites (STMS) markers, TA18 and TA180 (3.9 cM apart) were identified that flank the resistance gene. These findings could be the starting point for a Marker-Assisted Selection (MAS) programme for rust resistance in chickpea.     

Molecular characterization of a powdery mildew resistance gene in wheat cultivar suwon 92

ABSTRACT Powdery mildew, caused by Blumeria graminis f. sp tritici, is an important foliar disease of wheat worldwide. Pyramiding race-specific genes into a single cultivar and combining race-specific resistance genes with durable resistance genes are the preferred strategies to improve the durability of powdery mildew resistance. The objectives of this study were to characterize a powdery mildew resistance gene in Suwon 92 and identify gene-specific or tightly linked molecular markers for marker-assisted selection (MAS). A population of recombinant inbred lines (RILs) was derived by single seed descent from a cross between Suwon 92 and a susceptible cultivar, CI 13227. The RILs were screened for adult-plant infection type of powdery mildew and characterized with amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. The linked markers explained 41.3 to 69.2% of the phenotypic variances measured in 2 years. A morphological marker, hairy glume, was also associated with powdery mildew resistance in Suwon 92, and explained 43 to 51% of the phenotypic variance. The powdery mildew resistance gene in Suwon 92 was located on the short arm of chromosome 1A where Pm3 was located. Two gene-specific markers were developed based on the sequence of the cloned Pm3b gene. These two markers, which were mapped at the same locus in the peak region of the LOD score for the RIL population, explained most of the phenotypic variance for powdery mildew resistance in the RIL population. The powdery mildew resistance in Suwon 92 is most likely conditioned by the Pm3 locus. The gene markers developed herein can be directly used for MAS of some of the Pm3 alleles in breeding programs.     

结球甘蓝抗根肿病SSR分子连锁图谱的构建

结球甘蓝(Brassica oleracea L.var.capitata)简称甘蓝,是我国重要的蔬菜作物之一.甘蓝根肿病是由芸苔根肿菌(Plamodiophora brassicae Woron.)侵染引起的一种世界性真菌病害.近年来,该病在我国的发病面积急剧增加,造成甘蓝产量和品质大幅度降低,选育抗病品种成为防治根肿病的重要途径之一.     

Amplified fragment length polymorphism markers linked to a major quantitative trait locus controlling scab resistance in wheat

ABSTRACT Scab is a destructive disease of wheat. To accelerate development of scab-resistant wheat cultivars, molecular markers linked to scab resistance genes have been identified by using recombinant inbred lines (RILs) derived by single-seed descent from a cross between the resistant wheat cultivar Ning 7840 (resistant to spread of scab within the spike) and the susceptible cultivar Clark. In the greenhouse, F(5), F(6), F(7), and F(10) families were evaluated for resistance to spread of scab within a spike by injecting about 1,000 conidiospores of Fusarium graminearum into a central spikelet. Inoculated plants were kept in moist chambers for 3 days to promote initial infection and then transferred to greenhouse benches. Scab symptoms were evaluated four times (3, 9, 15, and 21 days after inoculation). The frequency distribution of scab severity indicated that resistance to spread of scab within a spike was controlled by a few major genes. DNA was isolated from both parents and F(9) plants of the 133 RILs. A total of 300 combinations of amplified fragment length polymorphism (AFLP) primers were screened for polymorphisms using bulked segregant analysis. Twenty pairs of primers revealed at least one polymorphic band between the two contrasting bulks. The segregation of each of these bands was evaluated in the 133 RILs. Eleven AFLP markers showed significant association with scab resistance, and an individual marker explained up to 53% of the total variation (R(2)). The markers with high R(2) values mapped to a single linkage group. By interval analysis, one major quantitative trait locus for scab resistance explaining up to 60% of the genetic variation for scab resistance was identified. Some of the AFLP markers may be useful in marker-assisted breeding to improve resistance to scab in wheat.     

Identification of a quantitative trait locus for high-temperature adult-plant resistance against Puccinia striiformis f. sp. hordei in 'Bancroft' barley

Sustainable control of plant diseases can be achieved by developing cultivars with durable resistance. 'Bancroft' barley has durable high-temperature, adult-plant (HTAP) resistance to stripe rust caused by Puccinia striiformis f. sp. hordei. The objectives of this study were to determine the inheritance of the HTAP resistance in Bancroft, develop molecular markers for the HTAP resistance using the resistance gene analog polymorphism (RGAP) technique, map the HTAP resistance quantitative trait locus or loci (QTL) on barley chromosomes, and determine the usefulness of the RGAP markers in other barley cultivars for marker-assisted selection. The parents and F(4) recombinant inbred lines (RIL) and the parents and F(5) RIL were evaluated in 2004 and 2005 in one and three field sites, respectively, in Washington State. Infection type (IT) and disease severity (DS) were recorded three times at each location during each growing season. Area under the disease progress curve (AUDPC) was calculated for each parent and RIL based on the DS data. Genetic analyses of IT data of the parents, F(1), and F(2) tested in the adult-plant stage under controlled high-temperature cycle in the greenhouse and the parents, F(4), and F(5) RIL in the field indicated that one dominant gene controlled the HTAP resistance in Bancroft. Using 119 F(5:6) RIL and IT data, a linkage map on chromosome arm 3HL was constructed with eight RGAP markers and three simple sequence repeat (SSR) markers. Using the QTL analysis, a QTL for HTAP resistance was mapped with the DS and AUDPC data on the same chromosome location as with the IT data. The QTL explained >70% of the total phenotypic variation for the DS and AUDPC. The heritability of the HTAP resistance based on the AUDPC data was 76%. The two markers most close to the QTL peak detected polymorphisms in 84 and 88% of 25 barley genotypes that do not have the Bancroft HTAP resistance when used individually, and detected polymorphism in 100% of the genotypes when used in combination, indicating that the markers could be used in incorporating the HTAP resistance into these barley genotypes to improve the level and durability of resistance to stripe rust.     

小麦抗白粉病基因Pm6的微卫星标记鉴定

 Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici, is a prevalent disease worldwide. Breeding and planting resistance cultivars have been proved effective and environmental friendly for control of the disease. To develop easily used PCR-based markers in marker assisted selection (MAS) for Pm6, a dominant powdery mildew resistance gene in wheat, 25 microsatellite markers on chromosome 2BL in wheat were screened between susceptible parent Yumai13 and resistance parent Timgalen carrying Pm6. F₂ population derived from Yumai13 and Timgalen was further analyzed by the marker Xgwm501. The results indicated that Xgwm501 was a co-dominant marker linked to Pm6 gene at a distance of 14.8 cM. 29 Pm-carrying varieties were tested by the marker Xgwm501 and only those carrying Pm6 showed 117 bp resistance specific band. This marker is proved to have high practicability and can be used in MAS of Pm6 gene in wheat breeding programs.