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1.
小麦几丁质酶基因Wch2的克隆与表达分析   总被引:4,自引:1,他引:3  
利用小麦几丁质酶基因PCR特异片段为探针,分离克隆了一个小麦Chidl几丁质酶基因Wch2。该基因编码311个氨基酸,不含内含子,具有一个信号肽、一个富含半胱氨酸的几丁质结合区域、两个变异区、两个酶活性区域。Southern分析表明,在小麦基因组中Wch2有多个拷贝。秆锈菌接种诱导Wch2在一对小麦近等基因系中差异表达;在抗病系中国春Srll中,接种3d后Wch2开始表达,6d后表达量更高;而在感病等基因系中国春srll中,在所有取样分析的时间内均未检测到Wch2表达。将Wch2克隆到细菌表达载体pET22b,在细菌中表达的重组Wch2具有几丁质酶活性。这些结果说明,分离的Wch2基因在小麦秆锈菌诱导的抗性反应中具有重要作用。  相似文献   

2.
本研究系统分析了大豆(品种:‘鲁豆4’)接种AM真菌Glomus fasciculatum和胞囊线虫(SCN,Heterodera glycines)4号生理小种后各处理菌根和线虫侵染率、几丁质酶和苯丙氨酸解氨酶(PAL)活性及几丁质酶基因Chib1和苯丙氨酸解氨酶基因PAL5转录物的动态变化。结果表明,接种SCN对AM真菌的侵染率没有产生显著影响,但先接种AM真菌后接种SCN的大豆根内线虫侵染率明显低于只接种SCN的处理。另外,先接种AM真菌后接种SCN的大豆根内几丁质酶和PAL活性显著提高,活性高峰出现在接种线虫后的第3天。值得注意的是,先接种AM真菌后接种SCN的大豆根内两种基因Chib1和PAL5转录物高峰也出现在接种SCN后的第3天,即AM真菌侵染率快速上升而SCN侵染率快速下降时期。所以Chib1和PAL5基因的表达可能是AM真菌诱导的抗大豆胞囊线虫病害防御反应的一种表现。因此推测Chib1和PAL5直接参与了AM真菌诱导大豆抗胞囊线虫病害的防御反应。  相似文献   

3.
本研究系统分析了大豆(品种:‘鲁豆4’)接种AM真菌Glomusfasciculatum和胞囊线虫(SCN,Heteroderaglycines)4号生理小种后各处理菌根和线虫侵染率、几丁质酶和苯丙氨酸解氨酶(PAL)活性及几丁质酶基因Chib1和苯丙氨酸解氨酶基因PAL5转录物的动态变化。结果表明,接种SCN对AM真菌的侵染率没有产生显著影响,但先接种AM真菌后接种SCN的大豆根内线虫侵染率明显低于只接种SCN的处理。另外,先接种AM真菌后接种SCN的大豆根内几丁质酶和PAL活性显著提高,活性高峰出现在接种线虫后的第3天。值得注意的是,先接种AM真菌后接种SCN的大豆根内两种基因Chib1和PAL5转录物高峰也出现在接种SCN后的第3天,即AM真菌侵染率快速上升而SCN侵染率快速下降时期。所以Chib1和PAL5基因的表达可能是AM真菌诱导的抗大豆胞囊线虫病害防御反应的一种表现。因此推测Chib1和PAL5直接参与了AM真菌诱导大豆抗胞囊线虫病害的防御反应。  相似文献   

4.
陈鹏  李振岐 《西北植物学报》2006,26(12):2468-2472
用0.4 mmol/L的苯并噻二唑(BTH)溶液处理小麦幼苗第1叶和第2叶2 d后接种白粉菌,比色法测定第3叶接种前后过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)、几丁质酶和β-1,3-葡聚糖酶的活性,结果表明BTH处理或接种均可使这4种酶活性升高。BTH诱导酶活性的系统增强与小麦对白粉病的诱导抗性密切相关。  相似文献   

5.
Shu W  Chen XH  Niu YC 《遗传》2011,33(9):1011-1016
为分析条锈菌诱导下的小麦抗病与感病近等基因系之间差异表达的基因,以接种小麦条锈菌CY26小种的抗病近等基因系Yr4/6×Taichung 29幼苗叶片cDNA作为实验方,接种CY26的感病亲本Taichung 29幼苗叶片cDNA为驱动方,利用抑制消减杂交(SSH)技术构建了一个包含1 300余克隆的消减文库。对文库中600个克隆进行了反向Northern点杂交筛选,对获得的阳性克隆进一步进行了Northern杂交验证,获得显著差异的克隆12个。经测序和BlastX分析,其中6个差异表达序列的推测产物分别为亮氨酸重复序列蛋白、过氧化氢酶、硫氧还蛋白、RNA结合蛋白、抗坏血酸过氧化物酶和热激蛋白。除亮氨酸重复序列为信号传导类蛋白外、其他几个均为抗病防御类蛋白。  相似文献   

6.
为了研究苯丙氨酸解氨酶基因与大蕉(Musa ABB cv. Dongguandajiao)抗枯萎病的关系,利用 RT-PCR 和 RACE技术克隆了大蕉苯丙氨酸解氨酶基因全长 cDNA。此 cDNA 长 1 300 bp,包含一个长为 1 191 bp,编码 397 个氨基酸的完整开放阅读框(ORF),推导的氨基酸序列与水稻 PAL 基因氨基酸序列同源性达 89%,将此基因命名为 M-PAL。Southern杂交结果表明大蕉中存在一个包含 4-5 个 PAL基因的基因家族,将此基因克隆到大肠杆菌表达载体 pET32(a )中,表达的蛋白质分子量大小与推导的相一致,并且表达的蛋白质表现出 PAL 酶活性。对接种香蕉枯萎病菌 4 号生理小种(Fusarium oxysporumf. sp. cubense (FOC) race 4 )后大蕉叶片中 M-PAL基因的转录谱进行研究表明,在接种枯萎病菌后,M-PAL基因在叶片中的转录水平提高,因此推测 M-PAL基因的表达可能与香蕉枯萎病抗性相关。  相似文献   

7.
丛枝菌根菌丝桥介导的番茄植株根系间抗病信号的传递   总被引:2,自引:0,他引:2  
菌根菌丝桥是植物间在地下进行物质交流的通道,但它能否作为植物间地下化学通讯的通道来传递抗病信号则缺乏研究.本文利用丛枝菌根真菌(AMF)摩西球囊霉在供体与受体番茄植株间建立菌丝桥,对供体植株接种早疫病病原菌茄链格孢菌,研究供体与受体番茄植株根系间是否存在抗病信号的传递.荧光定量PCR检测表明,AMF侵染后的供体番茄植株再接种病原菌,其根系中苯丙氨酸解氨酶基因(PAL)、脂氧合酶基因(LOX)和几丁质酶基因(PR3)的转录水平显著高于仅接种病原菌、未接种病原菌和AMF,以及只接种AMF的番茄植株.更重要的是,与供体有菌丝桥连接的受体番茄根系中PAL、LOX和PR3的基因的表达量也显著高于无菌丝桥连接、菌丝桥连接被阻断以及有菌丝桥连接但供体植物未接种病原菌的处理,3个基因最高转录水平达到无菌丝桥连接对照受体植物的4.2、4.5和3.5倍.此外,供体植株根系启动防御反应的时间(18和65 h)比受体(100和140 h)早.表明病原菌诱导番茄供体根系产生的抗病信号可以通过菌丝桥传递到受体根系.  相似文献   

8.
9.
麻疯树苯丙氨酸解氨酶启动子的克隆和表达载体的构建   总被引:2,自引:0,他引:2  
张淑文  高帆  秦小波  徐莺  陈放 《植物研究》2007,27(4):455-459
苯丙氨酸解氨酶(phenylalanine ammonia lyase, PAL)是苯丙烷类代谢途径的关键酶,催化苯丙氨酸转化为肉桂酸,促进黄酮、香豆素等次生代谢物的生成。本文根据已克隆的麻疯树苯丙氨酸解氨酶基因JcPAL的序列设计引物,通过DNA步移技术,克隆出长度为1 334 bp的JcPAL基因起始密码子上游序列。序列分析显示其不仅具备CAAT、TATA盒这些保守元件,而且包含多种胁迫诱导元件,特别是在序列中发现一些苯丙氨酸解氨酶特有的元件。为了鉴定JcPAL基因的启动子元件,分别将长度不同的5′端侧翼区缺失体定向插入载体pBI121中, 取代原有的CaMV35S启动子,构建了4个驱动报告基因GUS的植物表达载体。  相似文献   

10.
毛头鬼伞多糖对烟草酶活性和同工酶谱的影响   总被引:5,自引:0,他引:5  
分析了毛头鬼伞(Coprinus comatus)真菌多糖诱导烟草对烟草花叶病毒(TMV)抗性过氧化物酶(POD)、多酚氧化酶(PPO)、苯丙氨酸解氨酶(PAL)、几丁质酶、-β1,3-葡聚糖酶活性的变化。结果表明,毛头鬼伞多糖可提高POD、PPO、PAL、几丁质酶和-β1,3-葡聚糖酶的活性,接种TMV后毛头鬼伞多糖处理的烟草酶活性显著高于不处理者。上述结果提示,毛头鬼伞多糖处理后烟草酶活性的增强可能与其诱导烟草获得抗性有关。  相似文献   

11.
A glycopeptide elicitor prepared from germ tubes of the rust fungus Puccinia graminis Pers. f. sp. tritici Erikss. & Henn (Pgt), as well as chitin oligosaccharides, chitosan, and methyl jasmonate (MJ) stimulated lipoxygenase (LOX) activity (E.C. 1.13.11.12) in wheat (Triticum aestivum) leaves. Immunoblot analysis using anti-LOX antibodies revealed the induction of 92- and 103-kD LOX species after Pgt elicitor treatment. In contrast, MJ treatment led to a significant increase of a 100-kD LOX species, which was also detected at lower levels in control plants. The effects of chitin oligomers and chitosan resembled those caused by MJ. In conjunction with other observations the results suggest that separate reaction cascades exist, and that jasmonates may not be involved in Pgt elicitor action. LOX-92 appears to be mainly responsible for the increase in LOX activity after Pgt elicitor treatment because its appearance on western blots coincided with high LOX activity in distinct anion-exchange chromatography fractions. It is most active at pH 5.5 to 6.0, and product formation from linoleic and [alpha]-linolenic acid is clearly in favor of the 9-LOOHs. It is interesting that a 92-kD LOX species, which seems to correspond to the Pgt elicitor-induced LOX species, was also detected in rust-inoculated leaves.  相似文献   

12.
孔令让 《植物学报》2022,57(4):405-408
小麦条锈病是由条形柄锈菌小麦专化型(Puccinia striiformis f. sp. tritici, Pst)引起的真菌病害, 在全世界范围内危害小麦(Triticum aestivum)生产。培育和种植持久抗性小麦品种是控制小麦条锈病最有效的方法。由于病原体突变导致免疫受体逃避检测, 因此抗病基因经常失效。而易感基因(S基因)突变介导的抗性常具持久性与广谱性。近日, 西北农林科技大学植物免疫研究团队在揭示小麦受S基因保护的分子机制方面取得显著进展, 为抗病育种提供了有力工具。他们发现小麦感染条锈菌后, 真菌诱导受体样细胞质激酶TaPsIPK1与效应子PsSpg1特异性互作, 通过增强激酶活性和TaPsIPK1进入细胞核促进寄生。TaPsIPK1磷酸化转录因子TaCBF1d。TaCBF1d的磷酸化改变了其下游基因的转录活性。因此, TaPsIPK1和PsSpg1增强TaCBF1d磷酸化可能会重新编程靶基因表达, 干扰植物防御反应, 从而促进病原体感染。在2年的田间试验中, 小麦中TaPsIPK1的CRISPR-Cas9失活赋予了对Pst的广谱抗性, 且不影响重要的农艺性状。该研究首次揭示了由PsSpg1-TaPsIPK1-TaCBF1d在小麦条锈病S基因中触发的新的磷酸化转录调控机制, 为通过作物遗传修饰培育持久抗性品种提供了新策略。  相似文献   

13.

Key message

Fine mapping of the Ug99 effective stem rust resistance gene Sr45 introgressed into common wheat from the D -genome goatgrass Aegilops tauschii.

Abstract

Stem rust resistance gene Sr45, discovered in Aegilops tauschii, the progenitor of the D -genome of wheat, is effective against commercially important Puccinia graminis f. sp. tritici races prevalent in Australia, South Africa and the Ug99 race group. A synthetic hexaploid wheat (RL5406) generated by crossing Ae. tauschii accession RL5289 (carrying Sr45 and the leaf rust resistance gene Lr21) with a tetraploid experimental line ‘TetraCanthatch’ was previously used as the source in the transfer of these rust resistance genes to other hexaploid cultivars. Previous genetic studies on hexaploid wheats mapped Sr45 on the short arm of chromosome 1D with the following gene order: centromere–Sr45Sr33Lr21–telomere. To identify closely linked markers, we fine mapped the Sr45 region in a large mapping population generated by crossing CS1D5406 (disomic substitution line with chromosome 1D of RL5406 substituted for Chinese Spring 1D) with Chinese Spring. Closely linked markers based on 1DS-specific microsatellites, expressed sequence tags and AFLP were useful in the delineation of the Sr45 region. Sequences from an AFLP marker amplified a fragment that was linked with Sr45 at a distance of 0.39 cM. The fragment was located in a bacterial artificial chromosome clone of contig (ctg)2981 of the Ae. tauschii accession AL8/78 physical map. A PCR marker derived from clone MI221O11 of ctg2981 amplified 1DS-specific sequence that harboured an 18-bp indel polymorphism that specifically tagged the Sr45 carrying haplotype. This new Sr45 marker can be combined with a previously reported marker for Lr21, which will facilitate selecting Sr45 and Lr21 in breeding populations.  相似文献   

14.
Yang WY  Yu Y  Zhang Y  Hu XR  Wang Y  Zhou YC  Lu BR 《Hereditas》2003,139(1):49-55
Stripe rust is one of the most destructive diseases for wheat crops in China. Two stripe rust physiological strains, i.e. CYR30 (intern. name: 175E191) and CYR31 (intern. name: 293E175) have been the dominant and epidemic physiological strains since 1994. One Aegilops tauschii accession (SQ-214) from CIMMYT was found immune from or highly resistant to Chinese new stripe rust races CYR30 and CYR31 at adult stage. SQ-214 was crossed with a highly susceptible Ae. tauschii accession As-80. Analysis of data from F1-F2 populations of SQ-214/As-80 revealed that the resistance was controlled by a single dominant gene. To exploit the resistance for wheat breeding, SQ-214 was crossed with Chinese Spring (CS) and backcrossed by two Chinese commercial wheat varieties MY26 and SW3243. The resistance from SQ-214 was suppressed in the F1 hybrids (CS/SQ-214) and the F2 population of CS/SQ-214//MY26. However, the resistance of SQ-214 was expressed in several F2 individuals of CS/SQ-214//SW3243. Eleven advanced lines with high level of resistance to the Chinese stripe rust CYR30 and CYR31 have been developed. This result suggested that SW3243 does not suppress the expression of the Chinese stripe rust and should be used as wheat germplasm for exploiting resistance of Ae. tauschii in wheat breeding. The gliadin electrophoretic pattern of the eleven advanced lines with high stripe rust resistances was compared with their parents SQ-214, CS and SW3243 by acid polyacrylamide gel electrophoresis. The omega-gliadin bands of Gli-Dt1 in Ae. tauschii SQ-214 were transferred to some advanced lines and freely expressed in common wheat genetic background. One of advanced lines possesses a null Gli-D1 allele, where the omega-gliadin bands encoding by the Gli-D1 allele were absent. The potential utilization of this advanced line for wheat quality and stripe rust resistance breeding is also discussed in this paper.  相似文献   

15.
16.
Inheritance of leaf rust and stem rust resistance in 'Roblin' wheat.   总被引:2,自引:0,他引:2  
P L Dyck 《Génome》1993,36(2):289-293
The Canadian common wheat (Triticum aestivum L.) cultivar 'Roblin' is resistant to both leaf rust (Puccinia recondita Rob. ex. Desm.) and stem rust (Puccinia graminis Pers. f. sp. tritici Eriks. and E. Henn.). To study the genetics of this resistance, 'Roblin' was crossed with 'Thatcher', a leaf rust susceptible cultivar, and RL6071, a stem rust susceptible line. A set of F6 random lines was developed from each cross. The random lines and the parents were grown in a field rust nursery artificially inoculated with a mixture of P. recondita and P. graminis isolates and scored for rust reaction. The same material was tested with specific races of leaf rust and stem rust. These data indicated that 'Roblin' has Lr1, Lr10, Lr13, and Lr34 for resistance to P. recondita and Sr5, Sr9b, Sr11, and possibly Sr7a and Sr12 for resistance to P. graminis. In a 'Thatcher' background, the presence of Lr34 contributes to improve stem rust resistance, which appears also to occur in 'Roblin'.  相似文献   

17.
Wheat stem rust caused by Puccinia graminis f. sp. tritici can cause devastating yield losses in wheat. Over the past several decades, stem rust has been controlled worldwide through the use of genetic resistance. Stem rust race TTKSK (Ug99), first detected in Uganda in 1998, threatens global wheat production because of its unique virulence combination. As the majority of the currently grown cultivars and advanced breeding lines are susceptible to race TTKSK, sources of resistance need to be identified and characterized to facilitate their use in agriculture. South Dakota breeding line SD 1691 displayed resistance to race TTKSK in the international wheat stem rust nursery in Njoro, Kenya. Seedling screening of progeny derived from SD 1691 crossed to susceptible LMPG-6 indicated that a single resistance gene was present. Allelism and race-specificity tests indicated the stem rust resistance gene in SD 1691 was Sr28. The chromosome arm location of Sr28 was previously demonstrated to be 2BL. We identified molecular markers linked to Sr28 and validated this linkage in two additional populations. Common spring wheat cultivars in the central United States displayed allelic diversity for markers flanking Sr28. These markers could be used to select for Sr28 in breeding populations and for combining Sr28 with other stem rust resistance genes.  相似文献   

18.

Key message

Wheat lines carrying Ug99-effective stem rust resistance gene Sr43 on shortened alien chromosome segments were produced using chromosome engineering, and molecular markers linked to Sr43 were identified for marker-assisted selection.

Abstract

Stem rust resistance gene Sr43, transferred into common wheat (Triticum aestivum) from Thinopyrum ponticum, is an effective gene against stem rust Ug99 races. However, this gene has not been used in wheat breeding because it is located on a large Th. ponticum 7el2 chromosome segment, which also harbors genes for undesirable traits. The objective of this study was to eliminate excessive Th. ponticum chromatin surrounding Sr43 to make it usable in wheat breeding. The two original translocation lines KS10-2 and KS24-1 carrying Sr43 were first analyzed using simple sequence repeat (SSR) markers and florescent genomic in situ hybridization. Six SSR markers located on wheat chromosome arm 7DL were identified to be associated with the Th. ponticum chromatin in KS10-2 and KS24-1. The results confirmed that KS24-1 is a 7DS·7el2L Robertsonian translocation as previously reported. However, KS10-2, which was previously designated as a 7el2S·7el2L-7DL translocation, was identified as a 7DS-7el2S·7el2L translocation. To reduce the Th. ponticum chromatin carrying Sr43, a BC2F1 population (Chinese Spring//Chinese Spring ph1bph1b*2/KS10-2) containing ph1b-induced homoeologous recombinants was developed, tested with stem rust, and genotyped with the six SSR markers identified above. Two new wheat lines (RWG33 and RWG34) carrying Sr43 on shortened alien chromosome segments (about 17.5 and 13.7 % of the translocation chromosomes, respectively) were obtained, and two molecular markers linked to Sr43 in these lines were identified. The new wheat lines with Sr43 and the closely linked markers provide new resources for improving resistance to Ug99 and other races of stem rust in wheat.  相似文献   

19.
Ren Y  Li SR  Li J  Zhou Q  DU XY  Li TJ  Yang WY  Zheng YL 《遗传》2011,33(11):1263-1270
小麦条锈病是影响杂交小麦普及推广的重要因素。文章利用基因推导法和SSR分子标记技术,研究了温光型两系杂交小麦恢复系MR168的抗条锈性遗传规律及其控制基因染色体位置。结果表明,MR168对CY29、CY31、CY32、CY33等条锈菌生理小种表现高抗至免疫;对SY95-71/MR168杂交组合的正反交F1、BC1、F2和F3群体分单株接种鉴定显示,MR168对CY32号小种的抗性受1对显性核基因控制,该抗病基因来源于春小麦品种辽春10号。利用集群分离分析法(Bulked segregant analysis,BSA)和简单重复序列(Simple sequence repeat,SSR)分子标记分析抗病亲本MR168、感病亲本SY95-71及183个F2代单株,发现了与MR168抗条锈病基因连锁的5个微卫星标记Xgwm273、Xgwm18、Xbarc187、Xwmc269、Xwmc406,并将该基因初步定位在1BS着丝粒附近,暂命名为YrMR168;构建了包含YrMR168的SSR标记遗传图谱,距离YrMR168最近的两个微卫星位点是Xgwm18和Xbarc187,遗传距离分别为1.9 cM和2.4 cM,这两个微卫星标记可用于杂交小麦抗条锈病分子标记辅助育种。  相似文献   

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