小麦品种周8425B抗叶锈病基因LrZH84的EST标记

本研究利用周8425B与中国春杂交获得的F2和F3群体,采用分离群体分组分析法(BSA),获得了与LrZH84紧密连锁的EST分子标记BF482555,并在F2和F3群体中检测的连锁距离分别为3.8cM和4.5cM,该标记在抗感品种间扩增出了共显性的多态性片段。这些研究结果证明,通过EST建立小麦抗叶锈病基因的分子标记是可行的,同时为周8425B抗叶锈病基因的分子标记辅助育种奠定了基础。     

小麦品种周麦16的遗传构成分析

为解析小麦骨干品种周麦16的分子遗传学基础,本研究利用覆盖小麦全基因组的90 K SNP标记对小麦周麦16及其亲本进行全基因组扫描。结果表明,2个亲本对周麦16的遗传贡献差异较大,周8425 B对周麦16的遗传贡献率为64.32%,远高于周麦9号对周麦16的遗传贡献率(35.68%);在不同基因组水平上,周8425 B对周麦16的遗传贡献均高于周麦9号的遗传贡献率;在不同染色体上,2个亲本对周麦16的遗传贡献各有侧重,周麦9号在2 A、2 D、6 A、6 B染色体上对周麦16的遗传贡献率均达到60%以上,而在1 B、3 D、5 A、5 D和6 D染色体上周8425 B对亲本的遗传贡献率达到80%以上。本研究明确了周麦16的分子遗传构成,为周麦16利用提供参考依据。     

小麦条锈病抗性基因Yr5的序列标记位点和酶切扩增多态序列标记的构建

Yr5基因赋予了到目前为止美国所鉴定出的所有小麦条锈病病原小种的抗性。Yr5基因的共分离抗性基因类似多态(RGAP)标记是可以获得的，但是这些标记的应用需要有关聚丙烯酰胺凝胶电脉以及不同品种间可能不是多态性的条件。为了构建能在Yr5抗     

常见主栽小麦品种的抗条锈病评价

选取了河南地区主栽的30个小麦品种进行条锈病抗病性研究。试验结果表明,各小麦品种对条锈病抗性存在明显差异。参试品种中,许农7号对条锈病表现近免疫;许科718、泛麦8号、郑麦103、太空6号、矮抗58、百农4199、中麦175、豫教6号、许农5号、西农189、西农811、豫农949等12个品种对小麦条锈病表现高抗;华育198、秋乐2122、周麦27、百农418、横麦136、周麦22、偃细9433、豫农035、偃高21、周麦21、丰德存12号、百农207、运旱115等13个品种对小麦条锈病表现中抗;郑麦379、豫麦49、开麦20等3个小麦品种对小麦条锈病表现中感;遂选101表现高感。对小麦条锈病抗性的不同,有可能导致生产上小麦条锈病大流行,选育推广抗病品种有利于预防小麦条锈病的大面积发生。     

利用分子标记定位普通菜豆抗炭疽病基因

为了定位中国普通菜豆的抗炭疽病基因, 选取抗炭疽病地方品种红芸豆(国家库编号F2322)与高感菜豆品种京豆(国家库编号F0777)配制杂交组合, 构建F₂抗感分离群体和F_2:3家系, 用菜豆炭疽菌81号生理小种鉴定抗病性并分析遗传性。结果表明, 红芸豆对菜豆炭疽菌81号小种的抗性是由一显性单基因控制的, 暂将该基因命名为Co-F2322。用分离群体分组分析法(BSA)和SSR、CAPs分子标记技术, 将该基因定位在B1连锁群上, 利用软件Mapmaker 3.0和Mapchart 3.0计算标记与目的基因间的遗传距离, 检测到3个SSR标记BMc32、C871、Pvm98和2个CAPs标记g1224、g683与抗炭疽病基因连锁, 遗传距离分别为26.06、3.58、13.56、3.81和12.75 cM。     

“金手指”香蕉抗枯萎病基因同源序列的克隆与分析(英文)

本研究以抗镰刀菌枯萎病香蕉种质"金手指"(AAAB)为材料,根据已克降的抗枯萎病基因的NBS保守结构域设计简并引物,通过同源序列克隆获得了20条来自基因组DNA的RGA片段,大小为530 bp左右.根据其推断的氨基酸序列,经保守结构域分析,其结构域均为NB-ARC,属于non-TIR-NBS类候选抗病基因类序列.它们均具有P-loop(GMGGVGKTT),Kinase-2(LLVLDDIW),RNBS-B(CKVLFTTRS)及疏水氨基酸结构域GLPL(GLPLALKVL)等4个保守氨基酸基元.20个RGA之间核苷酸序列的相似性在41.1%～99.3%之间,氨基酸序列的相似性在33.2%～96.3%之间.同时,对分离得到的20条RGAs进行系统发育树分析,发现它们分布在5个不同的区域.并且所编码的氨基酸序列与已知抗枯萎病基因Fom-2、I2C-1、I2C-2和I2等编码的氨基酸序列表现出28%～54%的同源性,证明了抗病基因在进化上具有一定的保守性.因此,这些抗病基因同源片段(RGA)的分离将为进一步从香蕉中分离抗枯萎病基因打下基础,也可作为分子标记筛选香焦抗枯萎病的候选基因.     

小麦抗病基因同源序列(RGAs)的克隆与分析

RGA(抗性基因同源序列)法是克隆植物抗性基因的一种经济有效的方法,成为近年来的研究热点。本实验综合分析了拟南芥,西红柿,水稻,烟草等植物已克隆的抗性基因,并以这些抗性基因的NBS(核酸结合位点),LRR(富含亮氨酸重复),STK(丝氨酸/苏氨酸激酶)保守结构域设计并合成了几十对RGA引物,对小麦抗条锈病材料进行PCR扩增,获得以Xal-NBS为引物的R88RGA片段,经克隆和序列比对分析,发现该片段与逆境条件下植物抗病信号传导相关,与蛋白激酶同源性达到96%。此项研究对抗病机理的研究和基因的发掘有重要的指导意义。     

源于叙利亚小麦ICA31抗条锈病基因分析及分子标记研究

遗传分析表明,小麦材料ICA31携带一个显性抗条锈病基因,对流行的优势条锈菌小种条中30,31,32免疫;据等位性测定,ICA31抗条锈基因与已知抗锈基因Yr5、Yr10、Yr15不等位;从抗源的系谱分析,该基因来源于叙利亚普通小麦品系叙18;利用微卫星标记和分组分析(BSA)法,筛选到与该抗条锈病基因（Yr-Syria）紧密连锁的SSR标记WMS11-193;对F2分离群体142个单株分析结果表明,该抗条锈病基因（Yr-Syria）与WMS11-193间遗传距离为2.1cM;将Yr-Syria定位于小麦1BS上;为该基因进行抗条锈小麦分子辅助育种打下基础。     

普通小麦品种Brock抗白粉病基因分子标记定位

为明确利用Brock转育成的小麦抗白粉病品系3B529(京411*7//农大015/Brock, F₆)抗性的遗传基础,将高感白粉病小麦品系薛早和3B529杂交,获得F₁代、F₂分离群体和F_2:3家系。抗病性鉴定和遗传分析结果表明,3B529对E09小种的抗性受1对显性基因控制,暂被定名为MlBrock。利用BSA和分子标记分析,获得了与MlBrock连锁的3个SSR标记Xcfd81、Xcfd78、Xgwm159和2个SCAR标记SCAR203和SCAR112,根据SSR和SCAR标记在中国春缺体四体、双端体和缺失系的定位结果,将MlBrock定位在小麦染色体臂5DS Bin 0~0.63区间上。MlBrock与Xcfd81和SCAR203共分离,与SCAR112的遗传距离为0.5 cM。这些分子标记的建立有利于今后Brock抗白粉病基因分子标记辅助选择和基因聚合。综合抗白粉病基因MlBrock的染色体定位和抗谱分析结果,推测MlBrock很可能是Pm2基因。     

以SSR标记对普通菜豆抗炭疽病基因定位

由菜豆炭疽菌引起的菜豆炭疽病是危害我国菜豆生产的主要病害之一, 鉴定和发掘新的抗病基因对于菜豆抗病育种具有十分重要的意义。以来自安第斯基因库的我国菜豆抗炭疽病地方品种红花芸豆与感病地方品种京豆杂交的F₂群体为试验材料, 通过人工接种菜豆炭疽菌81号小种进行抗病性鉴定, 发现该分离群体中抗病植株数与感病植株数符合3∶1的分离比例, 确定红花芸豆对菜豆炭疽菌81号小种的抗性由显性单基因控制, 将此基因命名为Co-F2533。用分离群体分组分析法(BSA)和微卫星多态性分析(SSR)技术对红花芸豆中的抗炭疽病基因进行分子标记鉴定, 用Mapmaker3.0计算标记与目的基因间的遗传距离, 发现B6连锁群上的4个SSR标记BM170、Clon1429、BMD37、Clon410与抗炭疽病基因Co-F2533连锁, 遗传距离分别为6.6、18.4、20.9和30.9 cM, 这些SSR标记与Co-F2533基因在B6连锁群上的排列顺序为Clon1429-Co-F2533- BM170-BMD37-Clon410。根据基因所在连锁群的位置、抗病基因的基因库来源可知Co-F2533是一个新的来源于安第斯基因库的抗炭疽病基因。     

Molecular mapping of a stripe rust resistance gene in wheat cultivar Wuhan 2

Stripe rust (or yellow rust), caused by Puccinia striiformis f. sp. tritici, is one of the most destructive diseases of wheat worldwide. Growing resistant cultivars is the best approach to control the disease. To identify and map genes for stripe rust resistance in wheat cultivar ‘Wuhan 2', an F₂ population was developed from a cross between the cultivar and susceptible cultivar Mingxian 169. The parents, 179 F₂ plants and their derived F_2:3 lines were evaluated for responses to Chinese races CYR30 and CYR31 of the pathogen in a greenhouse. A recessive gene for resistance was identified. DNA bulked segregant analysis was applied and resistance gene analog polymorphism (RGAP) and simple sequence repeat (SSR) techniques were used to identify molecular markers linked to the resistance gene. A genetic map consisting of five RGAP and six SSR markers was constructed. The recessive gene, designated Yrwh2, was located on the short arm of chromosome 3B and flanked by SSR markers Xwmc540 and Xgwm566 at 5.9 and 10.0 cM, respectively. The chromosomal location of the resistance gene and its close marker suggest that the locus is different from previously reported stripe rust resistance genes Yr30, QYr.ucw-3BS, Yrns-B1, YrRub and QYrex.wgp-3BL previously mapped to chromosome 3B. Yrwh2 and its closely linked markers are potentially useful for developing stripe rust resistance wheat cultivars if used in combination with other genes.     

利用Fhb1基因功能标记选择提高黄淮冬麦区小麦品种对赤霉病的抗性

赤霉病已上升为黄淮冬麦区的主要病害, 提高小麦品种对赤霉病的抗性成为该麦区主要的育种目标之一。宁麦9号、生选6号、建阳798、建阳84、苏麦3号和宁麦13均携带Fhb1基因, 对赤霉病表现中抗水平以上。本研究以这6个品种(系)为供体, 分别与高感赤霉病的周麦16矮败小麦近等基因系杂交和回交, 构建6个回交群体。利用Fhb1基因的KASP标记在回交后代中进行基因型分析, 分别选择携带和不携带Fhb1基因的可育株, 对后代株系进行单花滴注接种鉴定和田间病圃自然鉴定。回交后代携带Fhb1家系整体抗性达到中感, 比不携带Fhb1家系的平均病小穗数低4.2 (P < 0.01), 平均病情指数低4.0, 比轮回亲本周麦16的平均病小穗数和病情指数分别低8.1 (P < 0.01)和28.4 (P < 0.01)。不同供体品种(系)回交后代在赤霉病抗性上表现出明显差异, 以生选6号为供体的回交后代家系抗性表现最好。本研究表明, 利用Fhb1基因分子标记辅助选择技术能够有效地提高黄淮冬麦区小麦品种的赤霉病抗性水平。     

小麦穗发芽抗性的4个分子标记有效性检验

为筛选出适宜黄淮麦区和长江中下游麦区种植的抗穗发芽白粒小麦品种或种质资源,以36份黄淮麦区和长江中下游麦区的主要品种（系）及地方品种为研究对象,对已报道的4个与穗发芽抗性相关的分子标记：Vp1B3、Xgwm155、Xgwm269和Xbarc170进行有效性验证。测定参试材料种子萌发指数（GI）,并用上述4种标记进行PCR扩增,对扩增条带进行统计分析。结果表明,GI值显示,红粒品种（GI均值为5.1%）明显较白粒品种（GI均值为28.0%）低;4种标记扩增出的带型中仅Vp1B3的845 bp片段能有效地区分36份小麦品种(系);GI值筛选出6份抗穗发芽品种（系）中,其中3份为Vp1B3标记鉴定,可作为黄淮麦区和长江中下游麦区小麦穗发芽抗性育种中首选基因资源。     

Identification and molecular tagging of a stripe rust resistance gene in wheat line P81

Stripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is one of the most devastating diseases in common wheat ( Triticum aestivum L.). With the objective of identifying and tagging a new gene for resistance to stripe rust in wheat line P81, F₁, F₂ and F_2:3 populations from the cross 'Chuanmai 28'/P81 were inoculated with Chinese PST race CYR32 in greenhouse and field trials. P81 carried a single dominant gene for resistance (designated YrP81 ) to CYR32. Tests of allelism showed that YrP81 was different from Yr5 , Yr10 , Yr15 and Yr26 . Simple sequence repeat (SSR) and resistance gene-analogue polymorphism (RGAP) between the parents were used for genotyping the F₂ populations. YrP81 was closely linked to four SSR loci on chromosome 2BS with genetic distances of 18.3 cM ( Xwmc25 ), 1.8 cM ( Xgwm429 ), 4.1 cM ( Xwmc770 ) and 5.3 cM ( Xgwm148 ). Two RGAP markers RGA1 (NLRR/XLRR) and RGA2 (Pto kin4/NLRR-INV2) were also closely linked to YrP81 with genetic distances of 4.7 and 6.3 cM, respectively. The linkage map of YrP81 and molecular markers was established in the order Xwmc25 - RGA2 - RGA1 - Xgwm429 - YrP81 - Xwmc770 - Xgwm148 . Pedigree analysis, response patterns with Chinese PST races and associations with markers suggested that YrP81 is a novel stripe rust resistance gene. The PCR-based microsatellite and RGAP markers identified here could be applied in selection of YrP81 in wheat breeding.     

Analysis of durable resistance to stem rust in barley

Summary Since the mid-1940's, barley cultivars grown in the northern Great Plains of the USA and Canada have been resistant to stem rust caused byPuccinia graminis f. sp.tritici. This durable resistance is largely conferred by a single gene,Rpg1, derived from a single plant selection of the cultivar Wisconsin 37 and an unimproved Swiss cultivar. At the seedling stage, barley genotypes withRpg1 generally exhibit low mesothetic reactions at 16–20° C and slightly higher mesothetic reactions at 24–28° C to many stem rust pathotypes. This resistance is manifested by a low level of rust infection and mostly incompatible type uredia on adult plants.Rpg1 reacts in a pathotype-specific manner since some genotypes ofP. g. f. sp.tritici are virulent on cultivars carrying this gene in the field. Several factors may have contributed to the longevity of stem rust resistance in barley, a) since barley is planted early and matures early, it can sometimes escape damage from stem rust inoculum carried from the south; b) one or more minor genes may augment the level of resistance already provided byRpg1; c) the cultivation of resistant wheat cultivars and eradication of barberry have reduced the effective population size and number of potential new pathotypes ofP. g. f. sp.tritici, respectively; and d) virulent pathotypes ofP. g. f. sp.tritici andP. g. f. sp.secalis have not become established. This situation changed in 1989 when a virulent pathotype (Pgt-QCC) ofP. g. f. sp.tritici became widely distributed over the Great Plains. However,Rpg1 may still confer some degree of resistance to pathotype QCC because stem rust severities have been low to moderate and yield losses light on barley cultivars carrying the gene during the last four seasons (1989–1992). Several sources of incomplete resistance to pathotype QCC have been identified in barley. To facilitate the transfer of resistance genes from these sources into advanced breeding lines, molecular marker assisted selection is being employed.     

小麦品种周麦22抗叶锈病的QTL定位

小麦叶锈病（leaf rust）是对小麦危害最严重的真菌病害之一,原菌群体中新致病菌类型的不断出现导致部分抗叶锈病基因的抗性功能逐步丧失,不断发掘和研究利用新抗源基因、培育种植抗病品种是控制该病害最有效的方法。周麦22在田间成株期对叶锈病表现出良好的抗性,为解析周麦22成株期抗叶锈病的遗传基础,将周麦22与铭贤169杂交构建遗传群体,获得255个F_2:3家系群体,经2个年度的大田成株期抗叶锈病鉴定,并利用复合区间作图法对该群体的抗叶锈病QTL进行定位分析。结果显示,该群体成株期检测到2个抗叶锈病QTL位点,分别位于1BL和2BS染色体上,命名为QLr.hebau-1BL和QLr.hebau-2BS,分别解释9.62%~11.88%和16.89%~20.99%的表型变异,该位点对叶锈病抗性表现稳定,均来自抗病品种周麦22。初步的遗传定位结果显示,QLr.hebau-2BS可能为已知抗叶锈病基因LrZH22,而QLr.hebau-1BL是新的抗病QTL。     

Fhb1基因改良‘周麦’品种赤霉病抗性研究

为加快‘周麦’品种赤霉病抗性改良,利用当地主栽‘周麦’品种（系）‘周麦22号’、‘周麦32号’、‘周11550’为母本,‘宁麦9号’、‘生选6号’、‘扬麦21号’等长江中下游地区抗赤霉病材料为父本配制一系列杂交组合,经过选择获得621份F₃~F₆后代材料。在田间选用来源于江苏和河南两地不同的赤霉病菌株,通过单花滴注法接种小穗进行赤霉病抗性鉴定,同时利用抗赤霉病主效基因Fhb1紧密连锁的诊断性标记His-InDel对后代材料进行分子检测。结果表明,江苏的赤霉病菌株接种后代材料田间鉴定为高抗和中抗的占总数的23.9%,而河南的赤霉病菌株感染后代材料田间鉴定为高抗和中抗占比为35.1%,说明后代材料的赤霉病抗性比感病亲本有明显的改良,而且材料对江苏的赤霉菌菌株的抗性比河南的赤霉菌菌株的抗性低。分子检测结果显示,携带Fhb1和不携带Fhb1的材料之间赤霉病抗性差异极显著。这表明利用Fhb1基因分子标记辅助选择技术可用于改良‘周麦’品种的赤霉病抗性。     

Identification and chromosomal locations of novel genes for resistance to powdery mildew and stripe rust in a wheat line 101-3

Wheat powdery mildew and stripe rust, caused by Blumeria graminis f.sp.tritici (syn. Erysiphe graminis f.sp.tritici) and Puccinia striiformis Westend., respectively, are two important fungal diseases of wheat in many regions in the world that cause significant annual yield losses. In the present study, a dominant powdery mildew and a dominant stripe rust resistance gene in wheat line 101-3 which derived from the progenies of the wide cross between common wheat and Dasypyrum villosum Candary L., was located on chromosome 6B and 1B, respectively, by monosomic analyses. The two genes are different from known resistance genes on chromosome 6B for powdery mildew and 1B for stripe rusts, suggesting that the two genes might be novel resistance genes for powdery mildew and stripe rust, respectively. It is uncertain whether the two genes are allelic or lined with other resistance genes located on chromosome 6B for powdery mildew and 1B for stripe rust. Further allelism tests are necessary to determine the relationships between the resistance gene and other genes located on chromosome 6B for powdery mildew and 1B for stripe rust through molecular markers.     

小麦新品种“山农20”抗病基因的分子检测

山农20是2011年和2012年分别通过国家黄淮南、北片审定的小麦高产多抗新品种,在国家区试抗病性鉴定和生产中都表现出良好的抗黄淮麦区主要病害的特性。本研究利用与小麦抗白粉病、条锈病、叶锈病、纹枯病基因和抗赤霉病主效QTL紧密连锁的SSR、SCAR、STS等标记对该品种进行了分子检测,发现山农20含有6个抗白粉病基因(Pm12、Pm24、Pm30、Pm31、Pm35和Pm36),6个抗条锈病基因(Yr5、Yr9、Yr15、Yr24、Yr26和YrTp1),2个抗叶锈病基因(Lr21和Lr26),1个抗纹枯病基因(Ses1),但未检测到抗赤霉病主效QTL。分子检测结果部分解释了山农20的优良抗病性,也为利用分子标记辅助选择培育抗病稳产小麦新品种提供参考。