小麦抗白粉病基因定位与分子标记

对小麦抗白粉病基因的遗传定位与分子标记进行了综述,介绍了小麦抗白粉病的遗传,并对今后的研究方向进行了讨论。     

小麦抗白粉病基因SSR标记定位

从波兰小麦与普通小麦感病品系‘中13’杂交后代中选育出小麦抗源材料WP6192,田间表现高抗白粉病,遗传分析表明其含有1对显性抗白粉病基因,暂定名为PmWP6192。用分离群体分组分析法筛选多态性SSR标记,并用F2代群体进行遗传连锁分析。结果表明,SSR标记Xgwm515、Xgwm249、Xgwm425、Xgwm372、Xg-wm630、Xbarc10、Xbarc220、Xbarc201和Xbarc353与PmWP6192基因连锁,相距最近的标记是Xbarc353,遗传距离为2.3cM。根据连锁标记所在的染色体位置,将PmWP6192定位于2AL染色体。通过基因来源分析和2AL染色体上已有抗白粉病基因的等位性分子检测,推断PmWP6192可能是1个新的抗白粉病基因。     

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.     

Transgenic Tobacco Expressing Zephyranthes candida Agglutinin Showing Enhanced Resistance to Aphids

Aphid is one of the most serious, sap‐sucking insect pests which cause significant losses of crop yields. The aim of this study is to investigate whether transgenic plants expressing Zephyranthes candida agglutinin (ZCA) could confer enhanced resistance to aphids. Tobacco was transformed with a plasmid, pCAMBIAZCA, containing the marker genes nptII and gusA and the Zephyranthes candida agglutinin gene (zca) via Agrobacterium tumefaciens‐mediated transformation. Twenty‐six independent transgenic plants were regenerated. Western blot analysis revealed ZCA expression at various levels in transgenic plants. Insect bioassay tests showed that transgenic plants expressing a high level of ZCA significantly inhibited the growth of the population of peach potato aphids (Myzus persicae Sulzer). This is the first report in which transgenic plants expressing ZCA conferred enhanced resistance to aphids. Our study suggests the zca gene could be a useful candidate for genetic engineering strategies in plants to provide aphid resistance.     

Transgenic Tobacco Expressing Pinellia ternata Agglutinin Confers Enhanced Resistance to Aphids

Tobacco leaf discs were transformed with a plasmid, pBIPTA, containing the selectable marker neomycin phosphotransferase gene (nptII) and Pinellia ternata agglutinin gene (pta) via Agrobacterium tumefaciens-mediated transformation. Thirty-two independent transgenic tobacco plants were regenerated. PCR and Southern blot analyses confirmed that the pta gene had integrated into the plant genome and northern blot analysis revealed transgene expression at various levels in transgenic plants. Genetic analysis confirmed Mendelian segregation of the transgene in T1 progeny. Insect bioassays showed that transgenic plants expressing PTA inhibited significantly the growth of peach potato aphid (Myzus persicae Sulzer). This is the first report that transgenic plants expressing pta confer enhanced resistance to aphids. Our study indicates that the pta gene can be used as a supplement to the snowdrop (Galanthus nivalis) lectin gene (gna) in the control of aphids, a sap-sucking insect pest causing significant yield losses of crops.     

转石蒜凝集素基因烟草的抗蚜虫性

利用农杆菌介导法将质粒pBILRA转化烟草(Nicotianatabacum L.),该质粒含有由花椰菜花叶病毒35S启动子(CaMV35S)引导的筛选基因新霉素磷酸转移酶基因(nptⅡ)及石蒜凝集素基因(lra).通过卡那霉素筛选获得了25株独立转基因烟草植株.Western blot分析表明,石蒜凝集素蛋白在不同转基因植株中表达量不同.对转基因T1代植株的遗传分析表明,lra基因在大多数独立转基因植株后代中以孟德尔3:1的分离比方式遗传.抗虫试验表明,表达较高水平石蒜凝集素蛋白的转基因烟草对桃蚜种群的生长具有明显的抑制作用.首次报道了表达石蒜凝集素基因的烟草对蚜虫具有抗性.石蒜凝集素基因可用于植物抗虫基因工程研究及应用.     

小麦白粉病抗性基因的聚合及其分子标记辅助选择

采用了在早代进行抗性鉴定、淘汰感病株、保留抗病株继续种植、较晚世代（F4代）进行抗性鉴定结合分子标记辅助选择的策略,提高了选到聚合抗性植株的效率。利用与Pm2、Pm4α、Pm8、Pm21紧密连锁或共分离的RFLP标记和PCR标记（SCAR标记）,对含有这些基因的优良品系间配制的杂交组合的F4代进行了分子标记辅助育种选择,并结合抗性鉴定,筛选到14株Pm4α Pm2I的植株,16株Pm2 Pm4α的植株,6株Pm8 Pm21的植株。应该引起注意的是,Pm2 Pm4α对混合白粉病菌的抗性达到高抗至免疫水平,而Pm2和Pm4α单独存在时抗性较差,表明聚合抗病基因植株的抗性提高了,为培育具有持久性抗性的品系或品种提供了新思路,它在实践和理论研究上都将具有重要意义。     

人工合成小麦抗白粉病未知基因的SSR标记

YAV-2/TEZ//A.SQ(895)是硬粒小麦与粗山羊草杂交获得的抗白粉病人工合成小麦。本研究利用人工合成小麦YAV-2/TEZ//A.SQ(895)与感白粉病的普通小麦品系品资50098杂交和自交获得的F2代群体及F3家系,在温室条件下鉴定群体的白粉病抗性。遗传分析结果表明,该抗白粉病基因为显性单基因遗传。利用647对小麦SSR引物进行了白粉病抗性基因的分子标记分析,结果表明该白粉病抗性基因与2A染色体的6个SSR标记连锁,与标记Xcfa2086的遗传距离最近,为11.8cM。     

Pathogenicity of the Fungus Verticillium lecanii to Aphids and Powdery Mildew

Aspects of the host specificity and pathogenicity of the hyphomycete, Verticillium lecanii , were investigated under laboratory conditions. DiVerences were observed in the pathogenicities of three strains of V. lecanii (DAOM 198499, DAOM 216596 and Vertalec) to the potato aphid Macrosiphum euphorbiae and Sphaerotheca fuliginea , the causal agent of cucumber powdery mildew. The estimated median lethal concentration required to achieve 50% mortality (LC ), 50 median lethal time leading to 50% mortality (LT ) and aphid net reproductive rate ( R ) 50 0 indicated that Vertalec and strain 198499 were more virulent to aphids than strain 216596. The estimated median colonization time for 50% of fungal colonies (CT ) showed that strain 50 198499 was the best antagonist of cucumber powdery mildew. Further comparison suggested that the mean pathogenicity of V. lecanii strain 198499 to cucumber powdery mildew was almost equivalent to that of Sporothrix flocculosa , a biological control agent of greenhouse fungal pathogens. These observations provide convincing experimental evidence that V. lecanii is biologically active against both arthropods and fungi. The potential of using V. lecanii strain 198499 in biological control is discussed.     

54份CIMMYT小麦材料抗白粉病分析

选用来自我国不同地区的20个白粉病菌毒性菌株,对54个CIMMYT小麦品种(系)进行抗病性分析.结果表明:(1)34个品种(系)含有抗病基因,以Pm8基因出现频率最高,有15个品种(系)携带该基因;(2)参试主效基因中,Pm1、Pm3e、Pm5、Pm6和Pm7基因已丧失对我国白粉菌的抗性,Pm16和Pm20基因的抗性最强;(3)50个1B/1R易位系品种(系)中31个含有抗病基因,48%的抗病1B/1R易位系可检测到Pm8基因.根据田间成株期病程曲线下面积(AUDPC)聚类分析结果,可将54份材料分为高抗、中抗、中感和高感4类,7个品种(系)不含任何主效抗病基因而田间表现中到高的抗性,是典型慢病性品种.     

小麦抗白粉病侵染初期的表达序列标签分析

以抗白粉病品系“百农 32 17×Mardler”　BC5F4为材料 ,构建了一个白粉病菌接种初期的抑制消减杂交cDNA文库 ,测序获得 76 0条ESTs。与GenBank序列进行BLASTx分析 ,获功能已知ESTs 2 71条。通过分析抗病相关基因 ,推测G蛋白介导的信号传导途径、SA信号传递系统、MAP相关信号传递系统等参与了小麦抗白粉病过程。SAR基因在抗病相关ESTs中的种类与数量最多。数据显示苯丙烷代谢途径、细胞壁结构修饰作用、细胞保卫机制参与了抗病过程。未知功能ESTs与GenBank序列进行BLASTn分析 ,其中许多与病原菌、非病原菌诱导cDNA文库来源的ESTs同源 ;新ESTs占全部ESTs的 16 6 %。     

On the Enhanced Callose Deposition in Barley with ml-o Powdery Mildew Resistance Genes

Carborundum treatment of barley leaves induced a callose deposition which was detected as diffuse blotches in the epidermal cells of susceptible barleys and as deeply stained tracks along the scratches in barleys with the ml-o powdery mildew resistance gene. Subsequent inoculation with powdery mildew resulted in appositions that enlarged inversely to their size in the respective varieties when inoculated without carborundum treatment. Aphids sucking the leaves resulted in rows of callose containing spots along the anticlinal cell walls. The spots were larger in the ml-o mutant than in the mother variety. Callose was deposited in connection with the pleiotropic necrotic spotting in barleys with the ml-o gene. Modification of the necrotic spotting by crossing the ml-o gene into other gene backgrounds did not result in any change in the size of appositions upon inoculation with powdery mildew. Callose deposition was never observed in cells subsidiary to the guard cells, and the absence of callose in such cells is suggested to be the reason for the sporadic occurrence of powdery mildew colonies on barleys with the ml-o gene.     

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.     

小麦新种质N9659抗白粉病基因的遗传分析

抗性种质N9659含有野生二粒小麦(资源编号:AS846)的抗白粉病基因,并对陕西关中地区白粉病优势小种表现为高抗.为了研究其抗白粉病基因的遗传规律,用高感品种辉县红、阿勃、陕160和阿勃21个缺(单)体系分别与N9659进行杂交,F1和F2苗期接种关中地区白粉病流行小种.结果显示,组合辉县红×N9659、阿勃×N9659、陕160×N9659及阿勃21个缺(单)体系和N9659杂交组合的所有F1均表现为高抗白粉病,F2的白粉病抗感比例除组合阿勃5BM×N9659偏离3∶ 1外,其他组合均符合3∶ 1,表明N9659苗期白粉病抗性由1对位于染色体5B上的完全显性基因控制.     

Genetics of Powdery Mildew Resistance in Barley

A comprehensive, critical review on the present knowledge regarding the genetics of resistance of barley to the powdery mildew fungus is presented. The review deals with six kinds of resistance: Race-specific resistance; Mlo resistance; partial resistance; induced resistance; passive resistance; and non-host resistance. Most of the sections are subdivided into: phenotype of the interaction; resistance mechanisms; and genetics. A distinction is made between three groups of genes involved in the defense of plants to diseases: those that serve exclusively to mediate resistance; those that are mobilized to strengthen the plants' defense; and those that serve exclusively functions other than disease defense, but may bring about resistance. The more than 200 gene symbols assigned to race-specific mildew resistance genes over time are summarized and revised to 85 symbols that may be considered valid.     

大麦DNA导入小麦产生抗白粉病变异的遗传研究

本研究将抗白粉病的大麦DNA通过花粉管途径直接导入感病的小麦品种花76中,后代出现13株抗白粉病变异株。其中5株在以后的世代中抗性稳定,另8株则继续分离。第2带分离株系的抗病株形成的第3代株系（或株行）中,抗性有分离的株行与无分离的株行比例为1.9:1,而分离株行内抗病株与不抗病株之比为3.35:1。抗性稳定株系与感病亲本杂交,F1表现高抗病,再与感病亲本回交,后代抗感病株比例为1:1,自交F2的比例为2.8:1。说明所获得的抗白粉病性受一对完全显性基因控制,抗病为显性。与已知抗白粉病基因的比较表明,这个抗病基因可能是来自大麦的一个新基因。13 Variant plants with immunity and high-resistance to powdery mildew were found in D1 generation from introducing resistant barley DNA into susceptible wheat cultivar, through pollen tube pathway after self pollination.Of the variants, 5 plants for the resistance had been stable and the other 8 plants segregated insuccessive generation.The ratio of segregating and stable plant-rows was 1.9:1 in D3 plant-rows derived from resistant plants of segregating D2-lines,and the ratio of resistant plants and susceptible plants was 3.35:1 among the segregating D3 plant-rows.The F1 -plants from crosses between stable resistant variants and susceptible parents were higgh resistant to powdery mildew.The ratio of resistant and susceptible plants was 1:1 in progenies of backcross of the F1 and susceptible parents, and this ratio was 2.8:1 in the F2 generation from the F1 selfing. Thus it can be seen that the resistance obtained is camtrolled by a pair of genes, the resistance is dominant. The results in comparison with known powdery mildew resistance genes in wheat indicated that the resistant gene obtained would be a new one from barley.     

大麦DNA导入小麦产生抗白粉病变异的遗传研究

本研究将抗白粉病的大麦DNA通过花粉管途径直接导入感病的小麦品种花76中,后代出现13株抗白粉病变异株。其中5株在以后的世代中抗性稳定,另8株则继续分离。第2带分离株系的抗病株形成的第3代株系（或株行）中,抗性有分离的株行与无分离的株行比例为1.9:1,而分离株行内抗病株与不抗病株之比为3.35:1。抗性稳定株系与感病亲本杂交,F1表现高抗病,再与感病亲本回交,后代抗感病株比例为1:1,自交F2的比例为2.8:1。说明所获得的抗白粉病性受一对完全显性基因控制,抗病为显性。与已知抗白粉病基因的比较表明,这个抗病基因可能是来自大麦的一个新基因。13 Variant plants with immunity and high-resistance to powdery mildew were found in D1 generation from introducing resistant barley DNA into susceptible wheat cultivar, through pollen tube pathway after self pollination.Of the variants, 5 plants for the resistance had been stable and the other 8 plants segregated insuccessive generation.The ratio of segregating and stable plant-rows was 1.9:1 in D3 plant-rows derived from resistant plants of segregating D2-lines,and the ratio of resistant plants and susceptible plants was 3.35:1 among the segregating D3 plant-rows.The F1 -plants from crosses between stable resistant variants and susceptible parents were higgh resistant to powdery mildew.The ratio of resistant and susceptible plants was 1:1 in progenies of backcross of the F1 and susceptible parents, and this ratio was 2.8:1 in the F2 generation from the F1 selfing. Thus it can be seen that the resistance obtained is camtrolled by a pair of genes, the resistance is dominant. The results in comparison with known powdery mildew resistance genes in wheat indicated that the resistant gene obtained would be a new one from barley.     

中国小麦白粉病发生地域分布的气候分区

根据全国１２０个气象站点小麦拔节一成熟期间的多年平均（１９７１—１９９８）旬≥０．１ｍｍ雨日出现机率、旬降雨量、旬平均相对湿度、旬平均温度、旬日照时效和白杨病病害资料,采用系统聚类分析法将中国小麦白杨病发生地域分布划分为发生气候带（１）和不发生气候带（ｌ）。Ｉ又分为常发气候亚带（１Ａ）、易发气候亚带（１Ｂ）、次易发气候亚带（１Ｃ）、偶发气候亚带（１Ｄ）。ＩＡ分为冬小麦白杨病西南常发气候区（１Ａ１）、冬小麦白杨病长江流域常发气候区（１Ａ２）、冬小麦白杨病淮河流域常发气候区（１Ａ３）共３个区５ＩＢ为冬小麦白杨病黄河流域易发气候区（１Ｂ区）１个区５ＩＣ分为南方冬小麦白杨病次易发气候区（１Ｃ１）、北方春小麦白杨病次易发气候区（１Ｃ２）、北疆春小麦白杨病次易发气候区（１Ｃ３）、西藏南部冬春小麦白杨病次易发气候区（１Ｃ４）共４个区５ＩＤ分为南方冬小麦白杨病偶发气候区（１Ｄ１）、北方春小麦白杨病偶发气候区（１Ｄ２）共２个区。评述了不同气候区小麦白杨病发生流行的气候生态特征。该分区结果客观揭示了小麦白杨病的发生地域分布规律,是针对不同气候条件下进行小麦白杨病分区预测预报和综合防治的基础性工作。     

农杆菌介导的大麦Mlo反义基因转化小麦获得抗白粉病后代

从大麦‘斯特林’幼叶总RNA中分离Mlo基因cDNA完整编码区,反向连接到植物双元载体（pBI-121.2）35S启动子下游,通过农杆菌介导的苗端转化法获得两种小麦基因型（‘烟优2801’和‘烟优361’）的转基因小麦。T0代405株中有55株PCR检测阳性,平均转化率达到13.58%,T0和T1基因组DNA Southern杂交可以证明大麦Mlo基因片段已整合到小麦基因组中并可传递到后代。两种基因型的转基因小麦T0和T1植株在温室及大田中均表现出对白粉病抗性的提高。农杆菌介导的苗端转化法可以简单、快速、高效地获得转基因株系;排除体细胞变异对转基因植株的影响;克服基因型对农杆菌转化的限制,是小麦遗传转化的一种实用方法。     

小麦抗白粉病基因Pm6的RAPD标记

从提莫菲维小麦转移到普通小麦中的小麦白粉病抗性基因Ｐｍ６是小麦白粉病（Ｅｒｙｓｉｐｈｅ ｈｒａｍｉｎｉｓ ｆ ｓｐ．ｔｒｉｔｉｃｉ）的有效抗性基因。用７００个随机引物对Ｐｍ６近等基因系进行ＲＡＰＤ分析,发现引物ＯＰＶ２０可在抗病近等基因系中产生大小为２ｋｂ的稳定的多态片段。用该引物检测１０个其他携Ｐｍ６的渐渗系材料,均可稳定扩增出该２ｋｂ的多态片段。理一步用ＯＰＶ２０对Ｐｍ６Ｆ２（ＩＧＶ１－４６３