光叶苕子(Vicia villosa var.)内生细菌Sz-2的特性及其对香蕉枯萎病的防治效果

【目的】从云南绿肥主推品种光叶紫花苕根内筛选出对香蕉具有促生和拮抗作用的菌株,并对其特性和香蕉枯萎病的防治效果进行研究,以丰富生防促生菌株资源库。【方法】从光叶紫花苕根中筛选出高效拮抗菌株Sz-2,使用平板对峙试验测定菌株抑菌效果;并测定其体外解磷、解钾、固氮及产铁载体能力;通过形态学观察、生理生化测定和16S rDNA序列分析,对该菌株进行了分类鉴定。布置盆栽试验,设4个处理:无抗生菌对照和接种病原菌TR4、Sz-2+TR4、Sz-2,测定香蕉叶片和球茎的发病指数以及促生效果。【结果】光叶紫花苕根内分离得到的菌株Sz-2对香蕉枯萎病菌具有较好的拮抗效果,平板径向抑菌率达74.3%。菌株Sz-2为革兰氏阴性细菌,菌落形态呈浅白色、放射状,接触酶、氧化酶、赖氨酸脱羧酶、β-半乳糖苷酶、精氨酸双水解酶、明胶水解、硝酸还原反应、纤维二糖、柠檬酸反应、葡萄糖发酵反应、酪素水解等均为阳性,吲哚反应、淀粉水解、产H2S反应、鸟氨酸脱羧酶、脲酶反应为阴性。结合形态学和分子生物学鉴定菌株Sz-2为嗜麦芽寡养单胞菌(Stenotrophomonas maltophilia)。盆栽结果显示,接种拮抗菌Sz-2后的香蕉球茎和叶片枯萎病发生较轻,球茎纵切面和叶片颜色正常且植株生长健康,对球茎和叶片的防效达69.66%和84.60%;香蕉茎粗、单株鲜重和株高与CK相比分别提高了6.01 mm、83.3 g和11.82 cm。【结论】菌株Sz-2对香蕉枯萎病具有较好的防治作用,加之其解磷、解钾和产铁载体功能,对香蕉植株具有显著的促生效果,可结合绿肥研制具有防控香蕉枯萎病的功能型绿肥,为香蕉促生抗病菌株的进一步田间开发应用提供理论依据。

Identification of Triticum aestivum-Haynaldia villosa Translocation Line T6BS·6BL-2VS

[Objective] The aim of experiment was to provide a new germplasm for wheat breeding by further using desirable genes in 2V chromosome of Haynaldia villosa.[Method] Through hybridization between common wheat(Triticum aestivum)-Haynaldia villosa disomic substitution line and common wheat Nonglin26-3C chromosome of Aegilops triuncialis disomic addition line,the analysis methods such as chromosome C-banding,genomic in situ hybridization and molecular marker technique were comprehensively applied and combined characters investigation.[Result] The wheat-Haynaldia villosa translocation line(T6BS·6BL-2VS)was selected from hybrid progenies to conduct characters investigation,which found some bristles on glume ridge of T6BS·6BL-2VS.[Conclusion] The translocation line induced by gametocidal chromosome was a small segment translocation line and the gene of bristle on glume ridge of Haynaldia villosa was located between the middle and the terminal of 2VS.

Microdissection of Haynaldia villosa Telosome 6VS and Cloning of Species-specific DNA Sequences

The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6VS was microdissected with a needle and transferred into a 0.5 mL Ep tube. In the 'single tube', all the subsequence steps were conducted. After two round of LA (Linker adaptor)_PCR amplification, the size of PCR bands ranged from 100 to 3 000 bp, with predominate bands 600-1 500 bp. The products were confirmed by Southern blotting analysis using Haynaldia villosa (L.) Schur. genomic DNA labeled with 32 P as probe. The PCR products were purified and ligated into clone vector-pGEM_T easy vector. Then, the plasmids were transformed into competence E. coli JM109 with cool CaCl 2. It was estimated that there were more than 17 000 white clones in the library. The size of insert fragments distributed from 100-1 500 bp, with average of 600 bp. Using H. villosa genomic DNA as probe, dot blotting results showed that 37% clones displayed strong and medium positive signals, and 63% clones had faint or no signals. It is demonstrated that there were about 37% repeat sequence clones and 67% single/unique sequence clones in the library. Eight H. villosa_specific clones were screened from the library, and two clones pHVMK22 and pHVMK134 were used for RFLP analysis and sequencing. Both of them were H. villosa specific clones. The pHVMK22 was a unique sequence clone, and the pHVMK134 was a repeat sequence clone. When the pHVMK22 was used as a probe for Southern hybridization, all the powdery mildew resistance materials showed a special band of 2 kb, while all the susceptible ones not. The pHVMK22 may be applied to detect the existence of Pm21.

Establishment of 6VS Telocentric Lines of Haynaldia villosa Resistant to Powdery Mildew Induced by Immature Embryo Culture

The line of T240-6 was selected from 32 SC 2 lines of immature embryo culture of T240 (common wheat (Triticum aestivum L.)× Wheat-Haynaldia villosa (L.) Schur. 6D/6V substitution line) through powdery mildew characterization, glutamate oxaloacetate transaminase (GOT) enzyme and gliadin (Gli) analyses and in situ hybridization. All of the individual plants resistant to powdery mildew lacked the locus of GOT at 6VL arm (GOT-V 2) and had gliadin locus at 6VS arm (Gli-V 2) of Haynaldia villosa. All the plants resistant to powdery mildew had one or two telocentric chromosomes that did not pair with wheat chromosomes but paired between themselves. T240-6 was identified as a telocentric line through in situ hybridization.

Production of Somatic Hybrid Plants Between Two Types of Wheat Protoplasts and the Protoplasts of Haynaldia villosa

小麦 (TriticumaestivumL .)济南 177的两种原生质体 ,一种来自快速生长的悬浮细胞 ,它们因长期继代而丧失分化能力 ,其染色体只有 2n =2 4～ 2 8;另一种来自可以再生的愈伤组织 ,其原生质体不能持续分裂。它们中任一种与UV照射过的簇毛麦原生质体融合均不能再生植株。然而当它们混合在一起作为受体时 ,能够获得再生绿色植株。细胞核基因和胞质基因的分析证明这些绿色植株是杂种。以上事实说明这两种原生质体在融合时存在某些互补的关系 ,讨论了这种融合方式的可能作用及重要性。

荒漠植物沙鞭(Psammochloa villosa)种质资源收集及谱系遗传分化初探

沙鞭(Psammochloa villosa)是禾本科(Poaceae)、针茅族(Stipeae)、沙鞭属(Psammochloa)中一个具有重要经济和生态价值的荒漠特有种,具有较强的抗逆性,是一种优质防风固沙植物,主要分布于内蒙古高原及其毗邻沙地。2013-2016年,本研究调查了沙鞭野生种质资源在内蒙古高原及其毗邻地区的自然地理分布,并在群体水平上收集了凭证标本、DNA材料和成熟种子;同时,从采集材料中选取具有代表性的180个个体进行了nrITS序列的扩增和测序,结果发现沙鞭所有参试种群的nrITS序列存在两个较为明显的谱系遗传分支,即一个分支主要由生长于内蒙古高原东部的所测个体组成,另一个分支由内蒙古高原西部的所测个体构成,这与依据外部形态特征和叶表皮微形态特征鉴分的结果相吻合。此外,本文还进一步讨论了这些丰富的种质资源对今后研究和利用沙鞭的重要性和相关途径。

两种南海倔海绵Dysidea villosa和Dysidea marshalla的化学成分

目的:从我国南海2种倔海绵属(Dysidea)海绵Dysidea villosa和Dysidea marshalla中提取分离具有生物活性的化学成分。方法:运用硅胶柱层析和凝胶柱层析分别对上述2种海绵的丙酮提取物进行分离纯化,根据其化学性质,结合现代波谱技术(MS,NMR等),对分离得到的化合物进行结构鉴定。结果:从海绵D.villosa中分离得到scalaradial(1),scalarin(2),12α-acetyl-19β,20α-dimethoxy-deoxascalarin(3),12α-acetyl-19α,20α-dimethoxy-de-oxascalarin(4),12-O-deaceyl-scalarin(5),(24R)-5α,8α-环二氧-麦角甾-6,22-二烯-3-醇(6),胆甾醇(7),squalene(8)和δ-selinene(9);从海绵D.marshalla中分离得到1～3,6～8。结论:化合物1对人白血病HL-60、人肝癌细胞BEL-7402及人乳腺癌MDA-MB-435等肿瘤细胞株表现出显著的抑制活性,其IC50分别为3.4,5.8和4.8μmol·L-1。

Development of EST-PCR Markers for the Chromosome 4V of Haynaldia villosa and Their Application in Identification of 4V Chromosome Structural Aberrants

EST-PCR based molecular markers specific for alien chromosomes are not only useful for the detection of the introgressed alien chromatin in the wheat background, but also provide evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers on chromosome 4V of Haynaldia villosa, a total of 607 primer pairs were designed according to the EST sequences, which were previously located in 23 different bins of wheat chromosomes 4A, 4B and 4D. By using the Triticum durum-H, villosa amphiploid and T. aestivum-H, villosa alien chromosome lines involving chromosome 4V, it was found that 9.23% of the tested primers could amplify specific bands for chromosome 4V. Thirty and twenty-six specific markers could be assigned to chromosome arms 4VS and 4VL, respectively. These 4V specific markers provided efficient tools for the characterization of structural variation involving the chromosome 4V as well as for the selection of useful genes located on chromosome 4V in breeding programs.

长绒毛栓菌(Trametes villosa DSM 9591)产漆酶的发酵条件优化

[目的]通过响应面法优化长绒毛栓菌发酵培养基,提高漆酶的产量。[方法]首先通过Plackett-Burman筛选出影响漆酶产量的3个主要因素;随后以葡萄糖浓度、二甲基苯胺浓度、摇床转速3因素自变量,采用Box-Behnken设计方法进行响应面试验,得到最佳发酵条件。[结果]回归分析得到最佳发酵条件方程,最佳的发酵条件为葡萄糖浓度12.37 g/L,二甲基苯胺浓度1.348 mmol/L,转速200r/min,此时漆酶的产量最高,经验证达4.1 U/ml,比初始条件提高了1.56倍。在10 L的发酵罐中,在1.0 VVM、350 r/min条件下培养时,漆酶产量最高可达13.6 U/ml。[结论]该研究得到了长绒毛栓菌产漆酶最佳的培养条件,为漆酶生产提供了更好的方法,为其今后的工业化生产奠定了基础。

Genetic Analysis and Molecular Mapping of an All-Stage Stripe Rust Resistance Gene in Triticum aestivum-Haynaldia villosa Translocation Line V3

Triticum aestivum-Hayaldia villosa translocation line V3 has shown effective all-stage resistance to the seven dominant pathotypes of Puccinia striiforms f.sp.tritici prevalent in China.To elucidate the genetic basis of the resistance,the segregating populations were developed from the cross between V3 and susceptible genotype Mingxian 169,seedlings of the parents and F 2 progeny were tested with six prevalent pathotypes,including CYR29,CYR31,CYR32-6,CYR33,Sun11-4,and Sun11-11,F 1 plants and F 3 lines were also inoculated with Sun11-11 to confirm the result further.The genetic studied results showed that the resistance of V3 against CYR29 was conferred by two dominant genes,independently,one dominant gene and one recessive gene conferring independently or a single dominant gene to confer resistance to CYR31,two complementary dominant genes conferring resistance to both CYR32-6 and Sun11-4,two independently dominant genes or three dominant genes（two of the genes show cumulative effect） conferring resistance to CYR33,a single dominant gene for resistance to Sun11-11.Resistance gene analog polymorphism（RGAP） and simple-sequence repeat（SSR） techniques were used to identify molecular markers linked to the single dominant gene（temporarily designated as YrV3） for resistance to Sun11-11.A linkage map of 2 RGAP and 7 SSR markers was constructed for the dominant gene using data from 221 F 2 plants and their derived F 2：3 lines tested with Sun11-11 in the greenhouse.Amplification of the complete set of nulli-tetrasomic lines of Chinese Spring with a RGAP marker RG1 mapped the gene on the chromosome 1B,and then the linked 7 SSR markers located this gene on the long arm of chromosome 1B.The linkage map spanned a genetic distance of 25.0 cM,the SSR markers Xgwm124 and Xcfa2147 closely linked to YrV3 with genetic distances of 3.0 and 3.8 cM,respectively.Based on the linkage map,it concluded that the resistance gene YrV3 was located on chromosome arm 1BL.Given chromosomal location,the reaction patterns and pedigree analysis,YrV3 should be a novel gene for resistance to stripe rust in wheat.These closely linked markers should be useful in stacking genes from different sources for wheat breeding and diversification of resistance genes against stripe rust.

Development of oligonucleotide probes for FISH karyotyping in Haynaldia villosa,a wild relative of common wheat

Haynaldia villosa is a wild relative of wheat and a valuable gene resource for wheat improvement.Owing to the limited number of probes available for fluorescence in situ hybridization(FISH),the resolution at which the karyotype of H.villosa can be characterized is poor,hampering accurate characterization of small segmental alien introgressions.We designed ten oligonucleotide probes using tandem repeats in DNA sequences derived from the short arm of H.villosa chromosome 6 V(6 VS).FISH with seven of them resulted in clear signals on H.villosa chromosomes.Using these,we constructed FISH karyotypes for H.villosa using oligo-6 VS-1 and oligo-6 VS-35 oligonucleotides and characterized the distribution of the two probes in five different H.villosa accessions.The new FISH probes can efficiently characterize H.villosa introgressions into wheat.

Heterologous expression of the Haynaldia villosa pattern-recognition receptor CERK1-V in wheat increases resistance to three fungal diseases

Wheat production is under continuous threat by various fungal pathogens.Identification of multipledisease resistance genes may lead to effective disease control via the development of cultivars with broad-spectrum resistance.Plant Lysin-motif(LysM)-type pattern-recognition receptors,which elicit innate immunity by recognizing fungal pathogen associated molecular patterns such as chitin,are potential candidates for such resistance.In this study,we cloned a LysM receptor-like kinase gene,CERK1-V,from the diploid wheat relative Haynaldia villosa.CERK1-V expression was induced by chitin and Blumeria graminis f.sp.tritici,the causal agent of wheat powdery mildew.Heterologous overexpression of CERK1-V in wheat inhibited the development of three fungal pathogens,thereby increased resistance to powdery mildew,yellow rust,and Fusarium head blight.CERK1-V physically interacted with the wheat Lys M protein Ta CEBi Ps.CERK1-V/Ta CEBi Ps interaction promoted chitin recognition and activated chitin signal transduction in wheat.Transgenic plants with excessively high CERK1-V expression showed high resistance but abnormal plant growth,whereas plants with moderate expression level showed adequate resistance level with no marked impairment of plant growth.In transgenic lines,RNA-seq showed that gene expression involved in plant innate immunity was activated.Expression of genes involved in photosynthesis,ER stress and multiple phytohormone pathways was also activated.Optimized expression of CERK1-V in wheat can confer disease resistance without compromising growth or defense fitness.

Two New Dihydroflavanoids from Patrinia villosa Juss

Two new dihydroflavonoids were obtained from the traditional Chinese medicinal herb Patrinia villosa Juss. Their structures were elucidated as （2S）-5, 7, 2＇, 6＇-tetrahydroxy-6, 8-di （γ, γ-dimethylallyl） flavanone （1） and （2S）-5, 7, 2＇, 6＇-tetrahydroxy-6-1avandulylated flavanone （2） by spectroscopic methods including UV, IR, HR-TOF-MS, 1D NMR and 2D NMR techniques.

The Meiotic Behavior of an Alien Chromosome in Triticum aestivum-Haynaldia villosa Monosomic Addition Lines

By the combination of cytological analysis and using genomic in situ hybridization technique to identify an alien chromosome in wheat-Haynaldia villosa monosomic addition lines, we studied the meiotic behavior of the alien chromosome. The results indicated that the frequency of bivalent pairing was lower than the value expected in PMCs of two monosomic addition lines, the frequency of wheat chromosomes unpairing increased, and the wheat homologous chromosome pairing was interfered with by the added chromosome 6V at metaphase I. The chromosome 6V lagged in 20.3% -29.3% of PMCs, sister chromatids 6V early divided in 29.0% - 34.1% of PMCs, the single chromosome 6V in 18.2% - 26.1% of PMCs went to a pole randomly, the breakage frequency of chromosome 6V was 1.2% - 2.9%. Meanwhile, it was also found that several wheat chromosomes showed earlier division, lagging and breakage in a few PMCs. It revealed that the added chromosome 6V influenced the behavior of wheat chromosomes at anaphase. It was also found that the translo-cation was produced between 6V and wheat chromosomes in 1.2% of PMCs. It offered evidence for transloca-tion between wheat and Haynaldia villosa 6V chromosomes.

Structural Changes of 2V Chromosome of Haynaldia villosa Induced by Gametocidal Chromosome 3C of Aegilops triuncialis

Haynaldia villosa （2n=2X= 14, VV）, a relative of wheat, plays important roles in wheat improvement mainly owing to its disease resistance. Powdery mildew resistance gene Pm21 has been successfully transferred into wheat by Cytogenetic Institute, Nanjing Agricultural University, China, and is widely used in the current wheat breeding programs. In this research, our objective is to further transfer and utilize the beneficial genes such as eye-spot resistance, yellow rust resistance, and gene of the tufted bristles on the glume ridge （a remarkable morphology） mapped on 2V of Haynaldia villosa. A disomic addition line with gametocidal chromosome 3C ofAegilops triuncialis added in Norin-26 was crossed to the wheat-H, villosa disomic substitution 2V（2D） and the hybrid F1 was then self-crossed. Chromosome C-banding, genomic in situ hybridization （GISH）, and meiotic analysis in combination with molecular markers were applied to detect the chromosome variations derived from hybrids Fz and F3. To date, four translocations including one small segmental translocation T6BS·6BL-2VS, two whole arm translocations （preliminarily designed as T3DS·2VL and T2VS.7DL） and one intercalary translocation T2VS·2VL-W-2VL, one deletion Del. 2VS·2VL-, one monotelosomic Mt2VS, and one isochromosome 2VS·2VS line have been developed and characterized. One wheat SSR marker Xwmc25.120 tagging 2VS and one wheat STS marker NAU/STSBCD135-1 （2BL） tagging 2VL were successfully used to confirm the alien chromosome segments involved in the seven lines. The tufted bristles on the glume ridge appeared in lines T2VS-7DL, Mt2VS, 2VS-2VS as well as the parent DS2V（2D）, whereas in T3DS·2VL, this trait did not appear. The gene controlling the tufted bristles was located on 2VS. Gametocidal chromosome 3C ofAegilops triuncialis could successfully induce chromosome 2V structural changes.

Two new bioactive salsolanol and biphenylsalsinol from the aerial parts of Salsola villosa Delile. ex Schul.(Chenopodiaceae) growing in Saudi Arabia

Objective: To isolate and characterize the bioactive secondary metabolites from aerial parts of widespread Chenopodiaceae taxa growing in Saudi Arabia: Salsola villosa Delile. ex Schul. Methods: Antibacterial activities of chloroformic extract, fractions and isolate compounds was evaluated against five bacterial strains(Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus and Staphylococcus epidermidis), using a paper disc diffusion method. The purification of compound(s) of chloroform extract was done by chromatographic column of silica gel. The structure elucidation was determined by extensive spectroscopic analysis(1H and 13 C nuclear magnetic resonance, correlation spectroscopy, heteronuclear multiple bond correlation, heteronuclear multiple quantum coherence and nuclear overhauser enhancement spectroscopy) and high resolution electrospray ionization mass spectroscopy analysis.Results: Bioactivity guided fractionation of the chloroformic extract led to the isolation of two bioactive compounds: 4-(4'-hydroxy-2'-methylcyclopent-2'-enyloxy)-4-methylcyclopent-2-enol(1) named salsolanol and 4'-[3-(hydroxymethyl)oxiran-2-yl]-3-[(E)-3-hydroxyprop-1-en-1-yl]-6, 2'-dimethoxy [1, 1'-biphenyl]-2-ol(2) named biphenylsalsinol. The antibacterial effects of the chloroform extracts, fractions and isolated compounds 1 and 2 were also evaluated in this work. Results showed that the compounds 1 and 2 exhibited antibacterial activities against four strains: Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Pseudomonas aeruginosa with diameter of zone of inhibition ranging between(9.33 ± 0.94) to(26.33 ± 0.94) mm.Conclusions: Based on data presented here, two new natural compounds secondary cyclic alcohol 1 and biphenylpropanoid 2 isolated from bioactive chloroformic extract from aerial parts of Salsola villosa can be responsible for its antibacterial activities.

Effect of Ferrum and Molybdenum Nutrition on Root Nodule and Physiological Growth of Vicia villosa Roth in Seedling Stage

[Objective] The paper was to explore the effects of ferrum and molybdenum nutrition on root nodule and physiological growth of Vicia villosa in seedling stage. [Method] A local cultivar of Qujing city,Vicia villosa,was studied using soilless cultivation method,and the effects of ferrum,molybdenum nutrition on root nodule and physiological growth of V. villosa in seedling stage were explored by adding 0. 01 mmol/L sodium molybdate and 0. 2 mmol/L ferrous sulfate respectively or simultaneously in the Hoagland's nutrient solution with the removal of ferrum and molybdenum salt. [Result]Ferrum and molybdenum significantly increased the number and weight of root nodule; the effect of single molybdenum treatment was the best,and the number and weight of root nodule in single molybdenum treatment were nearly 100% and 150% higher than that of the control. After treated by ferrum and molybdenum nutrition,the stem length and biomass of V. villosa decreased at varying levels,while root length and root shoot ratio increased significantly; the decrease extent of stem length in ferrum treatment and composite treatment was greater than that in molybdenum treatment,and the increase extent of root length in molybdenum treatment was the greatest. Application of ferrum and molybdenum fertilizer significantly improved ferrum and molybdenum content of V. villosa plants,and their soluble protein and free amino acid content increased at varying levels; single application of ferrum led to the greatest increase,while phosphorus content decreased at varying levels.The soluble sugar content of V. villosa decreased significantly at 90 d post ferrum and molybdenum application; application of molybdenum fertilizer reduced the starch content while ferrum fertilizer increased the starch content of V. villosa. Moreover,application of ferrum and molybdenum fertilizer reduced the SOD activity and enhanced the POD activity of V. villosa. [Conclusion] The application of ferrum and molybdenum fertilizer promotes the growth of root nodule and V. villosa plant.

Characterization of RAPD Markers,and the RFLP Marker Linked to Powdery Mildew Resistance Gene Derived from Different Accessions of H.villosa

The analysis was carried out on performance of the resistance gene from Haynaldia villosa accession of the former Soviet Union to different isolates of Bluemerie graminis. Polymorphisms were revealed between 6D/6V substitution line Pm930640 and its pedigree parents using five RAPD markers of OPAN031700, OPAI01700, OPAL03750, OPAD07480 and OPAG15580 screened out from 120 random 10-mers primers. Three RAPD markers of OPAN03, OPAI01 and OPAL03 were linked with the resistance gene by analysis of F2 population of Chancellor×Pm930640. Analysis of 29 wheat lines including part of lines conferring the known genes from Pm1 to Pm20 respectively, lines conferring resistance gene from two H. villosa accessions and the related wheat parents, were analyzed and the results showed that these markers not only linked to the gene resistant to powdery mildew from H. villosa, but also detected different genetic backgrounds. OPAL03750 can be used as the marker to distinguish the different resistant lines from two H. villosa accessions because it was only observed in the materials from H. villosa of the former Soviet Union. RFLP analysis also showed the polymorphisms between two H. villosa accessions and their derived resistant lines.

Molecular Characterization of a Triticum durum-Haynaldia villosa Amphiploid and Its Derivatives for Resistance to Gaeumannomyces graminis var. tritici

Take-all is a serious disease found in wheat across the world. Haynaldia villosa is considered to be resistant to take-all at a high level. TH3 was an amphiploid （2n =42, AABBVV） between Triticum durum and Haynaldia viUosa with significant resistance to take-all fungus isolated from China. In greenhouse experiment, the derivatives of the hybrid between wheat and TH3 showed better resistance to take-all than that of the wheat control. One of the derivatives named HW918-5 was selected for further analysis. Cytological and genomic in situ hybridization （GISH） analysis indicated that a monotelosome originated from H. villosa existed in the genome of the offspring of the line HW918-5. The monotelosome with promising resistant gene for take-all was located on the 3V chromosome of H. villosa in the further PCR-based molecular analysis.

Cloning and Characterization of a Family of Disease Resistance Gene Analogs from 6VS of Haynaldia villosa

In the present study, microdissection of 6VS and the cloning of the resistance gene analogs(RGA)from them were reported. The 6VS were microdissected with needle and 10 types of resistance gene analogs were obtained by PCR with degenerate oligonucleotide primer designed according to resistance genes. They were designated as Hvrgak1-Hvrgak10, GenBank accession numbers are AF387113-AF387121, AY040671- AY040672. Identity among RGAs was about 10-50%, and identity with cloned R gene from plants was 5-20%. Southern hybridization analysis results showed 3 RGAs, Hvrgak2, Hvrgak4, and Hvr-gak5 were linked with wheat powdery mildew resistance. These RGAs may be used as direct entrance or probes for cloning the disease resistance genes.