Molecular Cytogenetic Identification of a Wheat-Thinopyron intermedium （Host） Barkworth ＆ DR Dewey Partial Amphiploid Resistant to Powdery Mildew

Abstract: Wide cross and molecular cytogenetic methods were used to transfer the powdery mildew resistance gene from Thinopyron intermedium(Host) Barkworth & DR Dewey to wheat. Among the progeny of crossing common wheat (Triticum aestivum L.) Yannong 15 with Th. intermedium, a partial amphiploid E990256, with resistance to powdery mildew, was developed. It had 56 chromosomes and could form 28 bivalents in pollen mother cells at metaphase I of meiosis. Resistance verification by race 15 at the seedling stage and by mixed strains of Erysiphales gramnis DC. f. sp. tritici Em. Marchal at the adult stage showed it was immune to powdery mildew at both stages. Gene postulation via 21 isolates of E. gramnis f. sp. tritici and 29 differential hosts showed it was nearly immune to all the isolates used, and its resistance pattern was different from all the mildew resistance genes used, which indicated it probably contained a new resistance gene to powdery mildew. Biochemical verification showed it might convey different Th. intermedium chromosomes from those of the wheat‐ Th. intermedium partial amphiploids Zhong 1–5. Genomic in situ hybridization analysis by using St genomic DNA as the probe showed E990256 contained a recombination genome of St and E. (Managing editor: Wei WANG)     

兼抗白粉、条锈病小偃麦渗入系CH7124抗性遗传及细胞学鉴定

CH7124是通过八倍体小偃麦TAI8335与感病小麦杂交、回交育成的兼抗白粉病、条锈病的小偃麦种质系。利用抗性接种鉴定、细胞学和基因组原位杂交(GISH)技术相结合的方法,对CH7124的抗性来源、遗传方式及细胞学特征进行了分析和鉴定。结果表明,CH7124在苗期和成株期对条锈菌系CYR29、CYR31、CYR32、CYR33和白粉菌系E09、E20、E21、E26表现为免疫或近免疫,其抗性来自中间偃麦草,受1对显性核基因控制;CH7124的根尖细胞染色体数目为2n=42,花粉母细胞减数分裂中期I(PMC MI)绝大多数细胞内可观察到21个二价体,平均配对构型为2n=0.30 I+20.79 II+0.04 III;与普通小麦中国春、绵阳11的杂种F1中,有80%以上的花粉母细胞可观察到2n=21Ⅱ的染色体构型,其平均配对构型均为2n=21II。说明CH7124具有与普通小麦相似的染色体结构和规则的配对构型。由于利用以中间偃麦草总DNA为标记探针的原位杂交未观察到可见的外源DNA杂交信号,进一步证明CH7124是一个小麦-中间偃麦草的隐形异源渗入系。     

Expression of resistance to Blumeria graminis f.sp. tritici in 'Chinese Spring' wheat addition lines containing chromosomes from Hordeum vulgare and H. chilense

Blumeria graminis f.sp. tritici (syn. Erysiphe graminis f.sp. tritici) causes an important disease of wheat (powdery mildew) to which Hordeum vulgare and H. chilense are resistant. The study of chromosomal addition lines of H. vulgare and H. chilense in wheat showed that they possessed resistance to wheat powdery mildew. This was expressed as a reduction of disease severity but it was not associated with increased macroscopically visible necrosis. The resistance is of broad genetic basis, conferred by gene(s) present on different chromosomes of both H. vulgare and H. chilense. The feasibility of transferring this resistance to wheat is discussed.     

小偃麦衍生品系CH7086抗白粉基因的遗传及SSR分析

CH7086是兼抗白粉病、条锈病的小麦新品系,衍牛于来自十倍体长穗偃麦草的八倍体小偃麦与普通小麦的杂种后代.温室接种鉴定结果显示,CH7086对白粉病菌系E09、E21、E26均表现为免疫,且其抗件来自长穗偃麦草.抗性遗传分析表明CH7086的白粉病抗性由1对显性基因控制,暂定名为MlCH86.应用分离群体分组法(BSA)对从CH5241×CH7086的F2中随机选取的95个单株进行微卫星标记检测,发现位于2BL、2DL上的SSR位点Xbarc159在双亲和抗、感池间有特异性,并与抗性基因MlCH86连锁,其遗传距离为10.8 cM.用中国春第2部分同源群的缺体-四体系和双端体系进行验证,进一步将MlCH86定位在2BL上.用白粉病菌系E21、E26接种鉴定表明,MlCH86的抗性反应明显不同于2BL上已命名的抗性基因Pm6、Pm33.根据抗性基因的来源、染色体位置及抗性反应,初步推断存在于CH7086的抗性基因来自长穗偃麦草,它不同于已有的抗白粉病基因,可能是一个新基因.     

小偃麦附加系Z1和Z2中外源染色体2Ai-2的结构组成

小偃麦附加系Z1和Z2中外源染色体2Ai-2的结构组成@张增燕$中国农业科学院作物育种栽培研究所!北京100081@辛志勇$中国农业科学院作物育种栽培研究所!北京100081@陈孝$中国农业科学院作物育种栽培研究所!北京100081小偃麦;;附加系;;染色体     

普通小麦99-2439 中的白粉病抗性遗传

普通冬小麦品系99-2439在郑州连续4年对田间白粉菌(Blumeria graminis sp. tritici)表现高抗,但其抗性基因来源不清。通过染色体C-分带和1RS染色体特异性SCAR标记鉴定, 表明它是一个小麦-黑麦(Triticum aestivum - Secale cereale)1BL/1RS异易位系。通过对中国春×99-2439杂交F2代分离群 体抗性鉴定和1RS染色体臂检测结果分析, 证明该抗病基因不在1RS染色体臂上。用单孢小麦白粉菌分离株对其抗性遗传进行研究, 结果表明, 99-2439的白粉病抗性由一对小种专化、隐性抗病基因控制。由于携带Pm5a的Hope/8Cc对中国的21个小麦白粉菌分离菌株均高度感病, 而99-2439高抗混和白粉菌和5个单孢分离菌株, 所以, 99-2439所携带的抗白粉病基因不同于Pm5a。     

硬粒小麦-粗山羊草双二倍体白粉病抗性的遗传分析与基因推导

对99份硬粒小麦-粗山羊双二倍体用北京地区流行的5号白粉菌生理小种进行了白粉病抗性鉴定,筛选出11个苗期抗病的双二倍体材料和2个全生育期抗病的材料M53和M81。对M53和M81及其硬粒小麦和粗山羊草亲本进行的抗白粉病鉴定结果表明,其抗性来源于粗山羊草。与M53和M81具有相同硬粒小麦亲本、不同粗山羊草亲本双二倍体的抗性结果也表明抗性基因来源于粗山羊草。对M53和M81的抗性遗传分析表明,它们均携带1个单显性抗病基因。用14个白粉菌生理小种对已知抗病基因品系与M53和M81两份待测材料进行接种鉴定,结果表明,M53和M81与已知基因的抗菌谱均不相同,M53与M81的抗菌谱也不相同,说明M53和M81各自分别携带1个新的显性抗白粉病基因。     

普通小麦99-2439中的白粉病抗性遗传

普通冬小麦品系99-2439在郑州连续4年对田间白粉菌(Blumeria graminis sp.tritici)表现高抗,但其抗性基因来源不清.通过染色体C-分带和IRS染色体特异性SCAR标记鉴定,表明它是一个小麦-黑麦(Triticum aestivum-Secale cereale)lBL/1RS异易位系.通过对中国春×99-2439杂交F2代分离群体抗性鉴定和1RS染色体臂检测结果分析,证明该抗病基因不在1RS染色体臂上.用单孢小麦白粉菌分离株对其抗性遗传进行研究,结果表明,99-2439的白粉病抗性由一对小种专化、隐性抗病基因控制.由于携带Pm5a的Hope/8Cc对中国的21个小麦白粉菌分离菌株均高度感病,而99-2439高抗混和白粉菌和5个单孢分离菌株,所以,99-2439所携带的抗白粉病基因不同于Pm5a.     

Characterization of a partial wheat-Thinopyrum intermedium amphiploid and its reaction to fungal diseases of wheat

Partial amphiploids between wheat (Triticum aestivum L.) and Thinopyrum species play an important role in the transfer and use of traits from alien species. A wheat-Thinopyrum intermedium partial amphiploid, TAI8335, and its alien parent were characterized by a combination of genomic in situ hybridization (GISH) and cytological observations. Evidence from GISH indicated that the donor parent Th. intermedium possessed seven pairs of S, seven J(s) and 21 J chromosomes. Mitotic observation showed that the majority of TAI8335 plants had 56 chromosomes, but a few had 54 to 55, in some cases with two to three additional telochromosomes. The chromosomes in most pollen mother cells of plants with 2n = 56 formed 28 bivalents, averaging 27.12 in 223 cells, suggesting a basic cytological stability. Sequential GISH patterns using genomic Pseudoroegneria spicata and genomic Th. intermedium DNA as probes revealed that TAI8335 had fourteen chromosomes derived from Th. intermedium and its alien genome consisted of one pair of S-, three pairs of J(s) - and one pair of J-genome chromosomes as well as two translocated chromosome pairs, one being a Robertsonian translocation and another an intercalary translocation, both of which involved J and S genome. Two of the telochromosomes in the aneuploid plants originated from the J genome and one from wheat. Disease screening demonstrated this line was highly resistant to leaf rust, stem rust, stripe rust and powdery mildew. This study showed that the partial amphiploid TAI8335 appears to serve as a novel source for the transfer of resistance genes for multiple fungal pathogens to wheat.     

Powdery mildew resistance gene <Emphasis Type="Italic">Pm22</Emphasis> in cultivar Virest is a member of the complex <Emphasis Type="Italic">Pm1</Emphasis> locus in common wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L. em Thell.)

The powdery mildew resistance gene Pm22, identified in the Italian wheat cultivar Virest and originally assigned to wheat chromosome 1D, was mapped to chromosome 7A with the aid of molecular markers. Mapping of common AFLP and SSR markers in two wheat crosses segregating for Pm22 and Pm1c, respectively, indicated that Pm22 is a member of the complex Pm1 locus. Pm22 also showed a pattern of resistance reaction to a differential set of Blumeria graminis f. sp. tritici isolates that was distinguishable from those from other Pm1 alleles in lines Axminster/8*Cc ( Pm1a), MocZlatka ( Pm1b), Weihenstephan Stamm M1N ( Pm1c) and Triticum spelta var. duhamelianum TRI 2258 ( Pm1d). Based on these results, the gene symbol Pm1e is proposed for the powdery mildew resistance gene in cv. Virest.     

威岭栽培黑麦抗白粉病特性导入小麦的研究

威岭黑麦(Weiling rye)是一个高抗白粉病(Erysiphe gramininis f．sp．tritici)的中国矮杆栽培黑麦。以Weiling rye作为白粉病抗源,高感白粉病小麦栽培品种My8443为母本,从Weiling rye与小麦My8443远缘杂交的BC_2F_6后代中鉴定出一个新的小麦-黑麦易位系No．147,以实现威岭黑麦白粉病抗性向普通栽培小麦的转移。No．147及其亲本的抗白粉病特性通过苗期和成株期优势生理小种混合接种和室内单生理小种接种鉴定,改良的染色体C-分带和基因组原位杂交技术(GISH。Ge- nomic in situ hybridization)被用于鉴定小麦和黑麦的染色质,酸性聚丙烯酰胺凝胶电泳(APAGE)被用于鉴定黑麦醇溶蛋白1RS特异条带,11个黑麦种属特异性标记SCM(Secale cereale marker)引物被用于扩增分析黑麦特异性简单重复序列(SSR)。研究结果证实No．147是一个新的高抗白粉病的1BL/1RS小麦-黑麦染色体易位系,并对其产生的细胞学机制进行了分析。论文对中国栽培黑麦抗性基因资源的利用和该易位系在小麦遗传育种改良中的利用价值进行了讨论。     

Genome-wide association mapping for adult resistance to powdery mildew in common wheat

Molecular Biology Reports - Blumeria graminis f. sp. tritici, the causal agent of wheat powdery mildew disease, can occur at all stages of the crop and constantly threatens wheat production. To...     

Molecular cytogenetic identification of a wheat–<Emphasis Type="Italic">Aegilops geniculata</Emphasis> Roth 7M<Superscript>g</Superscript> disomic addition line with powdery mildew resistance

Aegilops geniculata Roth is an important germplasm resource for the transfer of beneficial genes into common wheat (Triticum aestivum L.). A new disomic addition line NA0973-5-4-1-2-9-1 was developed from the BC₁F₆ progeny of the cross wheat cv. Chinese Spring (CS)/Ae. geniculata SY159//CS. We characterized this new line by morphological and cytogenetic identification, analysis of functional molecular markers, genomic in situ hybridization (GISH), fluorescence in situ hybridization (FISH), and disease resistance evaluation. Cytological observations suggested that NA0973-5-4-1-2-9-1 contained 44 chromosomes and formed 22 bivalents at meiotic metaphase I. The GISH investigations showed that the line contained 42 wheat chromosomes and a pair of Ae. geniculata chromosomes. EST-STS multiple loci markers and PLUG (PCR-based landmark unique gene) markers confirmed that the introduced Ae. geniculata chromosomes belonged to homoeologous group 7. FISH identification suggested that NA0973-5-4-1-2-9-1 possessed an additional pair of 7M^g chromosomes, and at the same time, there were structural differences in a pair of 6D chromosomes between NA0973-5-4-1-2-9-1 and TA7661 (CS-AEGEN DA 7M^g). After inoculation with powdery mildew (Blumeria graminis f. sp. tritici, Bgt) isolates E09, NA0973-5-4-1-2-9-1 exhibited a powdery mildew resistance infection type different from that of TA7661, and we conclude that the powdery mildew resistance of NA0973-5-4-1-2-9-1 originated from its parent Ae. geniculata SY159. Therefore, NA0973-5-4-1-2-9-1 can be used as a donor source for introducing novel disease resistance genes into wheat during breeding programs with the assistance of molecular and cytogenetic markers.     

小麦硫代硫酸硫转移酶类似基因的克隆与定位

小麦-簇毛麦6VS/6AL易位系92R137含有抗白粉病基因Pm21。为了研究该易位系的抗病机理,应用mRNA差异显示和快速扩增cDNA未端（Rapid Amplification of cDNAEnd,RACE)技术对在白粉菌诱导后表达增强的基因进行了克隆,分离到1个命名为TaTST的全长cDNA序列。Northern杂交分析表明,TaTST基因在白粉菌诱导后表达明显增强,24h达到峰值,氨基酸序列同源性分析表明,TaTST与Datisca glomerata的硫代硫酸硫转移酶基因（rho-danese,EC,2.8.1.1)序列有64%相同,80%相似,用中国春缺体/四体系和端体系Southern杂交和基因特异性引物扩增（gene specific primer-PCR)将TaTST基因定位在小麦6B染色体短臂上,Southern杂交表明,该基因为单拷贝基因,由于在杨麦5号和6VS/6AL易位系间存在明显多态,可以推测在6VS上有TaTST的同源基因,TaTST是从小麦中分离的新基因。白粉菌诱导后的表达变化提示;TaTST与小麦抗白粉病反应有关。     

一粒小麦抗白粉病和条锈病基因的分析

一粒小麦是普通小麦抗性改良的宝贵资源.本研究对24份一粒小麦分别进行了白粉病和条锈病混合菌种苗期接种鉴定,进一步分别用一套白粉病菌菌株(15个)对2份乌拉尔图小麦和条锈病菌小种(21个)对1份栽培一粒小麦进行接种鉴定,其中乌拉尔图小麦UR206能抵抗所有供试白粉菌菌株,UR204除对白粉菌菌株E11感病外,对其余菌株表现抗性;栽培一粒小麦MO205对不同条锈菌小种表现出不同的抗性反应,研究表明乌拉尔图小麦UR206、UR204和栽培一粒小麦MO205分别含有与已知抗白粉病和抗条锈病基因不同的新基因.对乌拉尔图小麦UR204、UR206和栽培一粒小麦MO205分别进行抗白粉和条锈病基因的遗传分析,结果表明乌拉尔图小麦UR204和UR206分别含有一对显性抗白粉病基因,栽培一粒小麦MO205含有两对独立遗传的显性抗条锈病基因.     

St基因组中的CRW同源序列在偃麦草中的FISH分析

为了确定十倍体长穗偃麦草(Thinopyrum ponticum, Liu & Wang)和六倍体中间偃麦草(Th. intermedium, [Host] Barkworth & Dewey )的基因组组成, 根据野生一粒小麦(Triticum boeoticum)着丝粒自主型反转录转座子(CRW)序列设计特异引物, 以二倍体拟鹅观草(Pseudoroegneria spicata, Á Löve )基因组 DNA为模板进行PCR扩增, 筛选到一条St基因组着丝粒区相对特异反转录转座子的部分序列pStC1, 长度为1.755 kb (GenBank登录号: FJ952565), 其中有800 bp与小麦着丝粒反转录转座子(CRW)的LTR区高度同源, 另有小部分片段与其外壳蛋白编码基因(gag)部分同源, 并且包含一段富含AGCAAC碱基的重复序列。以pStC1为探针, 对十倍体长穗偃麦草的FISH检测结果显示其基因组组成为两个St组3个E组(St₁St₂E^eE^bE^x); pStC1与中间偃麦草杂交时, 不仅St基因组上有强烈的荧光信号, 而且E基因组一些染色体的近着丝粒区域也有杂交信号, 说明偃麦草属异源多倍体物种在其形成及进化过程中St与E基因组之间在着丝粒及近着丝粒相关区域可能存在协同进化。     

小麦Mlo及NBS-LRR类抗病基因同源序列的分离与鉴定

根据GenBank中公布的大麦白粉病抗性控制基因Mlo cDNA序列及一个来源于栽培一粒小麦(Triticum monococcum L.)的假定抗病基因序列分别设计引物,以携带小麦抗白粉病基因的近等基因系为材料进行RT-PCR筛选.结果获得两个表达基因的cDNA克隆.其中一个与大麦白粉病抗性控制基因Mlo的同源性达83%.另一个为非通读序列,含有两个可能的开放阅读框,分别包含抗病基因NBS保守结构域2和3以及与水稻抗稻瘟病基因Pib蛋白末端相似的13个LRR区域,推测该序列属于NBS-LRR类.白粉菌诱导前后,该片段RT-PCR扩增产物存在差异,表明该片段可能与小麦抗病性相关.利用"中国春"缺体-四体系,将该NBS-LRR类序列定位在小麦1D染色体上.     

用半定量RT-PCR方法分析小麦TaMlo-A1c基因的表达

以小麦稳定表达的肌动蛋白基因(Actin)作为对照,利用半定量反转录聚合酶链式反应(Semi-QRT-PCR)技术,对与抗白粉病有关的小麦(TriticumaestivumL.)TaMlo-A1c基因的表达进行了研究。结果发现:TaMlo-A1c基因在小麦的叶、茎、根中均表达,穗中不表达,在白粉菌(Blumeriagraminis(DC.)E.O.Speerf.sp.triticiEm.Marchal,Bgt)诱导不同时间后小麦叶片中的表达稍微有所增强。研究表明,用半定量RT-PCR技术研究小麦基因表达,具有特异性高、操作简便和可靠性强的优点。     

Genetic control of the wheat Triticum monococcum L. resistance to powdery mildew

Using hybrid analysis and test-clone method, 102 accessions of Triticum monococcum L. from the collection of the Vavilov All-Russia Institute of Plant Industry have been studied. This species of wheat has been found to by considerably polymorphic with respect to the resistance to the fungus Erysiphe graminis DC. f. sp. tritici Marchal. causing powdery mildew. The resistance of most accessions to the fungus population and clones is determined by dominant genes. In rare cases, the resistance was determined by recessive genes or one, two, or three oligogenes. A group of einkorn wheat accessions has been found in which the resistance to powdery mildew was determined by the same dominant factor or different but closely linked ones. Recessive resistance genes of T. monococcum differ from the recessive gene pm5 determining the resistance of T. aestivum plants. The genome of T. monococcum contains various genes of resistance to powdery mildew and is a potential source of effective genes to be used when selecting cultivated species of wheat for immunity.