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.     

水稻抗白叶枯病基因Xα-14在分子标记连锁图上的定位

选取对水稻白叶枯病原菌6个菲律宾小种均为感病的籼稻品种珍珠矮为母本,携带抗病基因Xα-14的粳稻近等基因系CBB14(抗菲律宾5号小种P5)为父本配制杂交组合。F1植株对相对应的菌系P5表现全生育期抗病, F2群体在分蘖前期接种鉴定结果表明,抗、感植株的分离符合3:1显性单基因分离比。根据对F2群体中选择的99个单株进行的RFLP分析的结果,构建了水稻第4连锁群的分子图谱,并把抗病基因Xα-14定位于RG62 0和G282之间。 Abstract:In order to map a rice bacterial blight resistance gene Xα-14,a F2 population containing 99 plants was generated from a cross between Zhenzhuai(susceptible to all six Philippines races)and CBB14 with Xα-14(resistant to race 5).It was observed that resistant and susceptible plants were segregated in a ratio of 3:1.Based on RFLP analysis in the population,a molecular linkage map of chromosome 4 was constructed,and the gene Xα-14 was located between RFLP markers RG 620 and G282.     

带有Ae.triuncialis抗白粉病基因的普通小麦-Ae.triuncialis单体附加系的培育

通过中国春Tal kr phlb基因综合体与Ae.triuncialis杂交、回交、自交及抗白粉病、细胞学鉴定,第一次获得了带有Ae.triuncialis抗白粉病基因的普通小麦-Ae.triuncialis单体附加系。 For the first time,the common wheat-Ae.trinucialis monosomic addition lines with resistant gene of Ae.triuncialis to powdery mildew were obtained by crosses and backcrosses between Chinese Spring Tal krph1b plants and Ae.triuncialis with Chinese Spring and identification of powdery mildew and cytology of their progenies.     

带有Ae.triuncialis抗白粉病基因的普通小麦-Ae.triuncialis单体附加系的培育

通过中国春Tal kr phlb基因综合体与Ae.triuncialis杂交、回交、自交及抗白粉病、细胞学鉴定,第一次获得了带有Ae.triuncialis抗白粉病基因的普通小麦-Ae.triuncialis单体附加系。 For the first time,the common wheat-Ae.trinucialis monosomic addition lines with resistant gene of Ae.triuncialis to powdery mildew were obtained by crosses and backcrosses between Chinese Spring Tal krph1b plants and Ae.triuncialis with Chinese Spring and identification of powdery mildew and cytology of their progenies.     

Cloning of a Resistance Gene Analog from Wheat and Development of a Codominant PCR Marker for Pm21

To Investigate the mechanism of resistance to wheat （Triticum aestivum L.） powdery mildew, suppression subtractlve hybridization was conducted between an isogenic resistant line carrying Pm21 and its recurrent parent Yangmal 5 to Isolate the resistance relative genes. A cDNA fragment specifically expressed in the resistant line was obtained and its full length was cloned by in silico cloning and RT-PCR. This gene encoded a deduced protein of 219 amino acids with a leucine-rich repeat （LRR） motif, often found In plant resistance genes, and was designated as Ta-LRR2. Ta-LRR2 had an increased expression level in the resistant line after Inoculation with Erysiphe graminis DC. f. sp. tritici Marchal. PCR analysis with different cytogenetlc stocks suggested that Ta-LRR2 was specifically associated with chromosome arms 6VS and 6AS. Linkage analysis further showed that Ta-LRR2 could be used as a resistance gene analog polymorphism marker of Pm21 for marker-assisted selection in germplasm enhancement and breeding practice. Moreover, how to Isolate Pm21 based on the Information obtained for Ta-LRR2 is discussed.     

Control of Rhyzopertha dominica in stored rough rice through a combination of diatomaceous earth and varietal resistance

Adults ofRhyzopertha dominica （F.）, the lesser grain borer, were exposed on four varieties of rough rice with Dobie indices of susceptibility of 1.1 to 1.1 （low）, and four varieties with Dobie indices of susceptibility of 3.4 to 3.8 （high）. The varieties with low and high Dobie indices were classified as resistant and susceptible, respectively, to R. dominica. The purpose of the study was to evaluate control of R. dominica through the use of diatomaceous earth （DE） in combination with rice varieties that were either susceptible or resistant to R. dominica. The rice was treated with varying rates of the commercial DE Insecto, up to a maximum of 1 000 mg DE/kg of rice. Adult mortality at each application rate of DE was generally greater on three of four resistant varieties compared to three of four susceptible varieties. Progeny production from the parental generation exposed on the rice was also greater in 3 of the 4 resistant varieties compared to 3 of the 4 susceptible varieties at DE rates of 500 mg/kg or more. Progeny production in rice treated with a maximum rate of 1 000 mg/kg DE ranged from 7-44 adults on the resistant varieties compared to 75-155 adults on the susceptible varieties. At DE rates of 500, 750, and 1 000 mg/kg, the percentage of insect-damaged kernels （IDK） was also greater in 3/4 resistant varieties than in the susceptible varieties. Results show combining the use of DE with varietal resistance of rough rice to R. dominica could be used to limit populations of this insect in stored rice and help prevent economic damage.     

Mapping of a Wheat Resistance Gene to Yellow Mosaic Disease by Amplified Fragment Length Polymorphism and Simple Sequence Repeat Markers

Wheat （Triticum aestivum L.） yellow mosaic virus （WYMV） is transmitted by a fungal vector through soil and causes serious wheat yield losses due to yellow mosaic disease, with yellow-streaked leaves and stunted plants. In the present study, the amplified fragment length polymorphisms （AFLP） and simple sequence repeat （SSR） were used to identify the molecular linkages with the resistance gene against WYMV. Bulked segregant analysis was performed with an F2 population derived from the cross of cultivar Ningmai 9 （resistant） × cultivar Yangmai 10 （susceptible）. By screening among the resistant or susceptible parents, the F2 pools and the individuals in the F2 population with 64 combined selective AFLP primers （EcoRI/MseI） or 290 reported SSR primers, a polymorphic DNA segment （approximately 120 bp） was amplified using the primer pair E2/M5, and an SSR marker （approximately 180 bp） was located on wheat chromosome 2A using the primer Xgwm328. Analysis with MAPMAKER/Exp Version 3.0b （Whitehead institute for Biomedical Research, Cambridge, MA, USA） indicated that these two markers were dominantly associated with the resistance gene at distances of 5.4 cM or 17.6 cM, respectively. The resistance gene to WYMV derived from Ningmai 9, is temporarily named YmNM, and was mapped to wheat chromosome 2A.     

Introgression of Resistance to Powdery Mildew Conferred by Chromosome 2R by Crossing Wheat Nullisomic 2D with Rye

Using the nulUsomic back-cross procedure, four wheat-rye chromosome substitution 2R （2D） lines with different agronomic performance, designated WR02-145-1, WR01-145-2, WR02-145-3, and WR02-145-4, were produced from a cross between 2D nullisomic wheat （Triticum aestivum L. cv. ＂Xiaoyan 6＂） and rye （Secale cereale L. cv. ＂German White＂）. The chromosomal constitution of 2n=42=21 in WR02-145 lines was confirmed by cytological and molecular cytogenetic methods. Using genomic in situ hybridization on root tip chromosome preparations, a pair of intact rye chromosomes was detected in the WR02-145 lines. PCR using chromosome-specific primers confirmed the presence of 2R chromosomes of rye in these wheat-rye lines, indicating that WR02o145 lines are disomic chromosome substitution lines 2R （2D）. The WR02-145 lines are resistant to the powdery mildew （Erysiphe graminis DC. f. sp. tritici E. Marchal） isolates prevalent in northern China and may possess gene（s） for resistance to powdery mildew, which differ from the previously identified Pm7gene located on chromosome 2RL. The newly developed ＂Xiaoyan 6＂- ＂German White＂ 2R （2D） chromosome substitution lines are genetically stable, show desirable agronomic traits, and are expected to be useful in wheat improvement.     

Seasonal changes of resistance to fenvalerate and cypermethrin in the larval parasitoid Cotesia plutellae （Hymenoptera： Braconidae）

The seasonal changes of insecticide resistance and stability in hymenopteran Cotesia plutellae, collected from Jianxin, Fuzhou-City, and Shangjie, Minhou-County, Fujian, China, were assessed by using a dry residual film method. The resistance to two insecticides in the field populations of C. plutellae was not stable under insecticide-free conditions in the insectarium. Compared with susceptible F11 progeny of C. plutellae in the insectarium, the resistance ratios （RR） in F0 parents were 18.4 for fenvalerate and 11.4 for cypermethrin based on LC50 at 9 hours, and 32.8 for fenvalerate and 28.5 for cypermethrin based on LC50 at 24 hours when the parasitoids were left in contact with the insecticides for 1 hour and mortalities were recorded at 9 and 24 hours, respectively. However, the RR in a field population of C. plutellae were 9.2 for fenvalerate and 12.7 for cypermethrin, if the parasitoids were left in contact with the insecticides for 24 hours. The resistances to the two pyrethroids in other field populations collected from Jianxin and Shangjie from November 2000 and July 2004 were also determined. Significant seasonal variations of resistance to the two insecticides in the field populations of C. plutellae were found. The RR were 3.0-18.4 for fenvalerate and 4.8-20.6 for cypermethrin in Jianxin populations from November 2000 to April 2002 based on LC50 at 9 h, and 2.3-13.6 for fenvalerate and 3.6-16.0 for cypermethrin in Shangjie populations from May 2002 to July 2004 based on LC50 at 24 hours. The resistance levels were high in spring and autumn and decreased sharply in summer. In addition, significant recovery from the knocked-down caused by the insecticides was found in the F0 and field populations of C. plutellae which were resistant to fenvalerate and cypermethrin if the parasitoids were left in contact with the pyrethroids for 1 hour. However, no recovery was found in susceptible F11 progeny.     

兼抗抗白粉病和黄矮病小麦育种材料的创造与鉴定

白粉病和黄矮病是小麦生产上的重要病害,近几年来这两种病害经常在我国一些小麦产区同时发生。为解决该问题,本研究通过杂交、回交方法将抗黄矮病的Bdv2基因（源自于YW642）和抗白粉病的Pm21基因（源自于CB037）聚合在一起,育成了兼抗黄矮病和白粉病的小麦新材料。通过田间抗病性鉴定与分子标记辅助选择相结合,得到聚合了Bdv2基因和Pm21基因的BC1代小麦22株,F2代小麦51株。农艺性状调查显示,这些含Pm21和Bdv2基因的双抗白粉病和黄矮病小麦新材料的农艺性状优于感病植株和原先的亲本,可以在小麦白粉病和黄矮病兼性抗病育种中作为优异种质资源加以利用。     

Transfer and mapping of a gene conferring later-growth-stage powdery mildew resistance in a tetraploid wheat accession

Wheat powdery mildew is a severe foliar disease and causes significant yield losses in epidemic years. Breeding and using resistant cultivars is the most widely employed strategy to curb this disease. To identify and transfer powdery mildew resistance genes in wild emmer wheat accession TA1410 into common wheat, a resistant F3 line derived from the cross of TA1410 × durum wheat line Zhongyin1320 was crossed with common wheat cultivar Yangmai158. The homozygous resistant BC5F2 lines derived from the backcross with Yangmai158 exhibited susceptibility at seedling stage and conferred increasing resistance when the plants were closer to heading stage. In two segregating BC5F3 families investigated at heading stage, the segregation of the resistance fit a 3:1 ratio, suggesting that a single dominant gene controls the resistance. This resistance gene, designated HSM1, was mapped to the 0.6-cM Xmag5825.1–Xgwm344 interval on chromosome 7AL and co-segregated with Xrga-C3 and Xrga-C6. A mapping position comparison with other powdery mildew resistance genes on this chromosome suggested that HSM1 belongs to the Pm1 resistance gene cluster. HSM1 is a useful candidate gene for resistance breeding, particularly in winter-wheat growing areas.     

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。     

TaRPP13-3, a CC-NBS-LRR-like gene located on chr 7D,promotes disease resistance to wheat powdery mildew in Brock

Disease resistance (R) gene, RPP13, plays an important role in the resistance of plants to pathogen infections; its function in resistance of wheat to powdery mildew remains unknown. In this study, a RNA-Seq technique was used to monitor expression of genes in susceptible wheat ‘Jing411’ and resistant near-isogenic line ‘BJ-1’ in response to powdery mildew infection. Overall, 413 differential expression genes were observed and identified as involved in disease resistance. RPP13 homologous gene on wheat chromosome 7D was preliminarily identified using the wheat 660K SNP chip. RPP13 was highly expressed in ‘BJ-1’ and encodes 1,027 amino acids, including CC, NB and LRR domain, termed TaRPP13-3. After inoculation with powdery mildew, expression of TaRPP13-3 in resistant wheat changed with time, but average expression was higher when compared to susceptible variety, thus indicating that TaRPP13-3 is involved in resistance to powdery mildew. Virus-induced gene silencing (VIGS) was used to inhibit expression of TaRPP13-3 in resistant parent ‘Brock’. Results indicated that silencing of TaRPP13-3 led to decreased disease resistance in ‘Brock’. Overall results of this study indicate that TaRPP13-3 gene is involved in the defence response of wheat to powdery mildew and plays a positive role in wheat powdery mildew interactions.     

来自西尔斯山羊草的抗小麦白粉病基因Pm57抗性丧失突变体的筛选与鉴定

小麦近缘种属来源的抗白粉病基因是培育小麦抗病品种,防治白粉病危害的最重要基因来源。Pm57是位于西尔斯山羊草2S^s#l染色体长臂上的一个外源基因,对小麦白粉病具有苗期和成株期广谱抗性。为了创制Pm57白粉病抗性丧失突变体,利用基于基因突变体的植物抗病基因克隆新兴技术分离Pm57基因,选用0.625%的甲基磺酸乙酯(EMS)对1万粒小麦-西尔斯山羊草Pm57易位系89(5)69种子进行了诱变处理,M1大田密播种植,收获了1598个M2可育株系。初步对其中300个M2株系进行苗期白粉病抗性接种鉴定,并利用2个Pm57基因特异分子标记X2L4g9P4/HaeⅢ和X284274及小麦全国区试品系DUS测试所用的42对SSR核心引物对Pm57抗性丧失突变体进行鉴定,筛选出来自27个M2株系的真实抗性丧失突变体70个,Pm57基因抗性丧失突变体频率达到9.0%。本研究所获得的白粉病抗性丧失突变体为Pm57基因的后续克隆与抗白粉病分子机理研究提供了重要的材料基础。     

Identification and mapping of a new powdery mildew resistance gene on chromosome 6D of common wheat

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most serious wheat diseases. The rapid evolution of the pathogen's virulence, due to the heavy use of resistance genes, necessitates the expansion of resistance gene diversity. The common wheat line D57 is highly resistant to powdery mildew. A genetic analysis using an F(2) population derived from the cross of D57 with the susceptible cultivar Yangmai 158 and the derived F(2:3) lines indicated that D57 carries two dominant powdery mildew resistance genes. Based on mapping information of polymorphic markers identified by bulk segregant analysis, these two genes were assigned to chromosomes 5DS and 6DS. Using the F(2:3) lines that segregated in a single-gene mode, closely linked PCR-based markers were identified for both genes, and their chromosome assignments were confirmed through linkage mapping. The gene on chromosome 5DS was flanked by Xgwm205 and Xmag6176, with a genetic distance of 8.3 cM and 2.8 cM, respectively. This gene was 3.3 cM from a locus mapped by the STS marker MAG6137, converted from the RFLP marker BCD1871, which was 3.5 cM from Pm2. An evaluation with 15 pathogen isolates indicated that this gene and Pm2 were similar in their resistance spectra. The gene on chromosome 6DS was flanked by co-segregating Xcfd80 and Xmag6139 on one side and Xmag6140 on the other, with a genetic distance of 0.7 cM and 2.7 cM, respectively. This is the first powdery mildew resistance gene identified on chromosome 6DS, and plants that carried this gene were highly resistant to all of the 15 tested pathogen isolates. This gene was designated Pm45. The new resistance gene in D57 could easily be transferred to elite cultivars due to its common wheat origin and the availability of closely linked molecular markers.     

Linkage of a RAPD marker with powdery mildew resistance

The aim of this study was to investigate the inheritance of powdery mildew disease and to tag it with a DNA marker to utilize for the marker-assisted selection (MAS) breeding program. The powdery mildew resistant genotype Fallon(er) and susceptible genotype 11760-3ER were selected from 177 genotypes by heavy infestation of germplasm with Erysiphe pisi through artificial inoculation. The F1 plants of the cross Fallon/11760-3 indicated the dominance of the susceptible allele, while F2 plants segregated in 3 : 1 ratio (susceptible : resistant) that fit for goodness of fitness by chi2 (P > 0.07), indicating monogenic recessive inheritance for powdery mildew resistance in Pisum sativum. A novel RAPD marker OPB18 (5'-CCACAGCAGT-3') was linked to the er-1 gene with 83% probability with a LOD score of 4.13, and was located at a distance of 11.2 cM from the er-1 gene.     

Effect of three suppressors on the expression of powdery mildew resistance genes in barley

Three recessive mutagen-induced alleles that partially suppress the phenotypic expression of the semidominant powdery mildew resistance gene Mla12 have been studied. When each suppressor is present in homozygous condition, the infection type 0, conferred by gene Mla12 when homozygous, is changed to intermediate infection types. The three suppressor lines were crossed with seven near-isogenic lines with different powdery mildew resistance genes and one, M100, was crossed with nine additional lines. Seedlings of parents and from the F1and F2 generations were tested with powdery mildew isolates that possessed the appropriate avirulence and virulence genes. The segregation of phenotypes in the F2 generation disclosed that the three suppressors affected the phenotypic expression of three resistance genes, whereas that of four resistance genes remained unaffected. The suppressor in mutant M100 affected the phenotypic expression of 9 of the 10 additional resistance genes present. It is suggested that the three suppressors are mutationally modified genes involved in host defence processes. This implies that different resistance genes employ different, but overlapping, spectra of defence processes, or signal transduction pathways. Key words : barley, Hordeum vulgare, powdery mildew, Erysiphe graminis hordei, mutation, resistance, suppressor.     

Genetic characterization of powdery mildew resistance in U.S. hard winter wheat

Powdery mildew significantly affects grain yield and end-use quality of winter wheat in the southern Great Plains. Employing resistance resources in locally adapted cultivars is the most effective means to control powdery mildew. Two types of powdery mildew resistance exist in wheat cultivars, i.e., qualitative and quantitative. Qualitative resistance is controlled by major genes, is race-specific, is not durable, and is effective in seedlings and in adult plants. Quantitative resistance is controlled by minor genes, is non-race-specific, is durable, and is predominantly effective in adult plants. In this study, we found that the segregation of powdery mildew resistance in a population of recombinant inbred lines developed from a cross between the susceptible cultivar Jagger and the resistant cultivar 2174 was controlled by a major QTL on the short arm of chromosome 1A and modified by four minor QTLs on chromosomes 1B, 3B, 4A, and 6D. The major QTL was mapped to the genomic region where the Pm3 gene resides. Using specific PCR markers for seven Pm3 alleles, 2174 was found to carry the Pm3a allele. Pm3a explained 61% of the total phenotypic variation in disease reaction observed among seedlings inoculated in the greenhouse and adult plants grown in the field and subjected to natural disease pressure. The resistant Pm3a allele was present among 4 of 31 cultivars currently being produced in the southern Great Plains. The genetic effects of several minor loci varied with different developmental stages and environments. Molecular markers associated with these genetic loci would facilitate incorporating genetic resistance to powdery mildew into improved winter wheat cultivars.     

A major QTL for powdery mildew resistance is stable over time and at two development stages in winter wheat

Despite the large impact of powdery mildew in wheat cultivated areas, little has been done to study powdery mildew resistance by QTL analysis up to now. The objective of the present paper is to present how the genetic basis of powdery mildew resistance in the resistant wheat line RE714 have been studied by QTL analysis at the adult plant stage over the course of 3 years, and at the vernalized seedling plant stage, and a comparison between the results obtained. Two segregating populations (DH and F_2:3) were derived from the cross between the resistant line (RE714), and a susceptible line (Hardi); these were analysed for powdery mildew resistance at the adult plant stage in the field under natural infection conditions in 1996, 1997 and 1998. The DH population was also tested for powdery mildew resistance at the vernalized seedling stage with four different isolates of powdery mildew. At the adult plant stage, a total of three QTLs (on chromosomes 5D, 4A and 6A) and five QTLs (on chromosomes 5D, 6A, 7A and 7B) were found for the DH and F_2:3 populations, respectively. The genetic control of resistance was found to be polygenic but involved a major QTL (on chromosome 5D), which was detected each year and which explained a high proportion of the variability observed (28.1%–37.9%). At the vernalized seedling stage, two QTLs were found (on chromosomes 5D and 7B) and the QTL detected on chromosome 5D was common to the four isolates tested. The comparison between the two development stages showed that the QTL on chromosome 5D was detected in all the different environments tested and again explained a high proportion of the variability. Different molecular interpretations of this QTL have also been discussed. Received: 5 October 2000 / Accepted: 1 March 2001