Effect of Two Herbicides on the Growth of Early Seedlings of Rye(Secale cereale)

[Objective]The aim was to study the effect of herbicide on the growth of early seedlings of rye（Secale cereale）.[Method]Effect of two kinds of herbicide（Atrazine and APM）on seedling growth of rye was investigated at the physiological,biochemical and cellular level.[Result]The Atrazin significantly decreased the contents of chlorophyll a,b and soluble proteins.Rye seeds were treated with 0.01-1 mg/L Atrazine for 16 h,the contents of chlorophyll a and b decreased from 1.26（a）,0.49（b）mg/g FW（control）to 1.15（a）,0.46（b）mg/g FW（0.1 mg/L）and 0.81（a）,0.33（b）mg/g FW（1.0 mg/L）.The content of soluble protein decreased with the increasing concentration of Atrazin.Atrazin had no significant influence on the cell division and chromosome structure variation.The contents of chlorophyll a,b and soluble proteins had no significantly change under the treatment of APM,but the number of chromosome structure variation such as chromosome bridge,multipolar division cells,lagging chromosome and unequal division cells increased significantly.[Conclusion]The critical concentration of Atrazine was 0.1-1.0 mg/L and 4 mg/L of APM in rye.

Development and identification of two novel wheat-rye 6R derivative lines with adult-plant resistance to powdery mildew and high-yielding potential

Powdery mildew,caused by Blumeria graminis f.sp.tritici(Bgt),is a devastating disease that seriously threatens wheat yield and quality.To control this disease,host resistance is the most effective measure.Compared with the resistance genes from common wheat,alien resistance genes can better withstand infection of this highly variable pathogen.Development of elite alien germplasm resources with powdery mildew resistance and other key breeding traits is an attractive strategy in wheat breeding.In this study,three wheat-rye germplasm lines YT4-1,YT4-2,and YT4-3 were developed through hybridization between octoploid triticale and common wheat,out of which the lines YT4-1 and YT4-2 conferred adult-plant resistance(APR)to powdery mildew while the line YT4-3 was susceptible to powdery mildew during all of its growth stages.Using genomic in situ hybridization,multi-color fluorescence in situ hybridization,multi-color GISH,and molecular marker analysis,YT4-1,YT4-2,and YT4-3 were shown to be cytogenetically stable wheat-rye 6R addition and T1RS.1BL translocation line,6RL ditelosomic addition and T1RS.1BL translocation line,and T1RS.1BL translocation line,respectively.Compared with previously reported wheat-rye derivative lines carrying chromosome 6R,YT4-1 and YT4-2 showed stable APR without undesirable pleiotropic effects on agronomic traits.Therefore,these novel wheat-rye 6R derivative lines are expected to be promising bridge resources in wheat disease breeding.

Molecular cytogenetic analyses of two new wheat-rye 6RL translocation lines with resistance to wheat powdery mildew

Rye(Secale cereale)is a valuable gene donor for wheat improvement,especially for its resistance to diseases.Developing rye-derived resistance sources is important for wheat breeding.In the present study,two wheat-rye derivatives,designated JS016 and JS110,were produced by crossing common wheat cultivar Yangmai 23 with Pakistani rye accession W2A.Using sequential genomic in situ hybridization(GISH)and multicolor fluorescence in situ hybridization(mc-FISH),JS016 and JS110 were identified as a T6BS.6RL translocation line and a T6BS.6BL6RL translocation line,respectively.Ten newly 6RL chromosome arm-specific markers were developed and used to confirm the 6RL translocation.The wheat 55K single-nucleotide polymorphism(SNP)array further verified the molecular cytogenetic identification results above and clarified their breakpoints at 430.9 and 523.0 Mb of chromosome 6B in JS016 and JS110,respectively.Resistance spectrum and allelism test demonstrated that JS016 and JS110 possessed novel powdery mildew resistance gene(s)that was derived from the 6RL translocation but differed from Pm20.Moreover,JS016 and JS110 had better agronomic traits than the previously reported 6RL translocation line carrying Pm20.To efficiently transfer and detect the 6RL translocation from JS016 and JS110,one 6RL-specific Kompetitive allele specific PCR(KASP)marker was developed and validated in high throughput marker-assisted selection(MAS).

小麦抗叶锈基因Lr45的SCAR标记

【目的】建立小麦抗叶锈基因Lr45SCAR(sequenced characterized amplified region序列特征扩增区域)标记。【方法】以小麦抗叶锈病基因材料TcLr45和感病材料Thatcher为亲本,利用黑麦基因组特异的RAPD标记引物OPH20进行PCR扩增,对获得的与预期大小相同的1.5kb的特异片段进行克隆测序,并根据该序列设计一对特异PCR引物LRYR和LRYF,对TcLr45×Thatcher F2代单株构建的分离群体进行扩增,验证该标记与Lr45的连锁关系,Mapmaker3.0软件绘制遗传连锁图。【结果】该标记在TcLr45中扩增出单一条带,片段大小为1272bp(命名为Ypsc20H1272),而在感病亲本中则无扩增条带。F2代分离群体进行连锁分析,遗传距离为8.2cM,该标记为与Lr45连锁的SCAR标记。【结论】将Lr45的RAPD标记转化成SCAR标记。

小麦-黑麦1RS/1BL新易位系的创制和分子细胞遗传学鉴定

利用普通小麦(Triticum aestivumL.)品种小偃6号与黑麦(Secale cerealeL.)品种德国白粒杂交,选育出一批带有黑麦抗病性状的小偃6号类型种质材料。应用连续C-分带-基因组原位杂交(sequent C-banding-GISH)技术对上述材料进行染色体组成分析,筛选出2个小麦-黑麦1RS/1BL纯合易位系BC152-1-1和BC01-89-1。其中,BC152-1-1(2n=42)除含有1对1RS/1BL易位染色体外,未见其他染色体变异;BC01-89-1(2n=43)除含有1对1RS/1BL纯合易位染色体外,还附加1条两端缺失的3R染色体。高分子量麦谷蛋白亚基(HMW-GS)组成分析和品质分析结果表明,BC152-1-1和BC01-89-1不仅含有来自小偃6号的14+15优质亚基,而且其蛋白质含量、湿面筋含量和SDS沉降值等品质性状都得到显著改良。

Primary Identification of Alien Chromatin in T911289,a Maintainer of Wheat Male Sterile Line with Cytoplasm of Aegilops kotschyi

The genomic composition of 1911289, a wheat ( Tritium aestivum L.) maintainer of K-CMS, was examined by several methods, such as genomic in situ hybridization (GISH), biochemical marking, and DNA molecular marking. The results got by GISH and PCR amplification of dispersed rye-specific repetitive DNA sequence suggested that the alien chromatin in T911289 derived from rye. Specifically PCR amplification of the rye-specific microsatellite primers (SCM9) and seed storage protein analysis indicated that the alien chromatin in T911289 had developed from the short arm of 1R chromosome of rye (1RS). PCR amplification by using microsatellite primers locating on 1BS and seed storage protein analysis also revealed that 1911289 had lost the arm of 1BS or a small distal segment of it. We conclude that T911289 is a heterogeneous population which displays two distinct different types of translocation, i.e. the Robertsonian translocation and small segment translocation. The Robertsonian translocation type observed in our study is different from the 1BL/1RS translocation which is widely used in wheat production; it may be a novel and complex translocation form. Though the linkage between the desirable agronomic traits and the deleterious genes expressed as sticky dough has not got broken in T911289, the recovery of small segment translocation will still benefit the genetic study of wheat and rye.

发酵谷物中产L-赖氨酸益生菌的筛选与鉴定

为了扩展高产L-赖氨酸菌种在食品、饲料中的应用与提高微生物发酵生产L-赖氨酸的安全性,从面包、麻花、馒头、发面饼、大列巴、黏豆包发酵谷物的生面团及米酒、白酒曲中分离培养出66株产L-赖氨酸益生菌菌株。将这些菌株进行培养、发酵,并利用高效液相色谱技术检测其发酵液中L-赖氨酸的含量,筛选出发酵液富含L-赖氨酸的菌种两株。最后利用16S rDNA分析及Blast分析、特异性引物PCR技术鉴定出这两株菌为肠膜明串球菌ATCC8293和德氏乳杆菌ATCC BAA-365。其发酵液中L-赖氨酸含量分别为52.33、46.09g/L,较其他菌种相对较高。

黑麦属NBS-LRR类抗病基因同源序列的分离与鉴定

根据已知植物抗病基因（resistance gene,Rgene）编码的蛋白质NBS-LRR保守结构设计了10对特异简并引物,对5份黑麦材料的基因组DNA进行抗病基因同源序列克隆,共获得45条抗病基因同源序列（resistance geneanalogs,RGAs）。同源性比较发现,RGAs序列之间的核苷酸相似性是41.9%~99.6%,其中16条具有连续开放阅读框（ORFs）。同时,利用MEGA 3.1将这16条序列与4个已知R基因进行聚类分析,发现有13条序列与RPM1亲缘关系近,2条与Pm3a亲缘关系近。本研究在黑麦中获得的RGAs既可用于筛选黑麦的抗病候选基因,同时也可以作为分子标记,用于作物分子辅助选择育种,且对进一步研究黑麦中NBS-LRR类R基因的起源和遗传进化提供参考和依据。

PEG-6000模拟干旱胁迫对冬黑麦生理指标的影响

为探究干旱胁迫对冬黑麦生长及生理指标的影响,使用15%聚乙二醇-6000(PEG-6000)对水培俄罗斯冬黑麦BK01模拟干旱胁迫,分别在处理0(CK)、3、6、9、12、24 h测定不同胁迫时叶片各项生理指标。结果表明,随胁迫时间的延长,冬黑麦叶片脯氨酸和丙二醛含量呈上升趋势;过氧化氢酶、过氧化物酶、超氧化物歧化酶活性、可溶性蛋白和可溶性糖含量呈先上升后下降趋势;相对含水率呈下降趋势;研究结果将为冬黑麦抗旱研究进展提供新的理论依据。

黑饲麦不同部位水浸提液的化感抑草作用研究

为探究饲用作物黑饲麦的化感作用,本试验以黑饲麦为供体植物,自生油菜和小麦为受体植物,采用培养皿生物测定法,分别测定黑饲麦的不同部位(根际土壤、根、秸秆、穗)、不同浓度(100 mg/mL、50 mg/mL、35 mg/mL、25 mg/mL)水浸提液对受体植物发芽率、芽长、根长、鲜重、干重的影响,并对其化感指数进行分析评价。结果表明,黑饲麦水浸提液对杂草自生油菜种子的萌发及幼苗生长存在化感抑制作用,其中秸秆、穗部抑制作用最显著,且随着质量浓度的升高,化感抑制作用增强,当质量浓度为100 mg/mL时,化感抑制作用最强;但黑饲麦水浸提液对小麦种子的萌发与生长并未表现出明显影响,因此,不会影响作物小麦的安全生长。

小麦-黑麦6RS/6AL易位染色体的遗传稳定性及其在配子中的传递

小麦-黑麦6RS/6AL易位系HM812-41携带抗白粉病基因Pm56。为评价其育种利用潜力,以HM812-41为亲本分别与推广品种蜀麦580、蜀麦830和蜀麦969杂交,杂种F1与中国春进行正反交,研究6RS/6AL易位染色体在不同背景中的遗传稳定性及其通过雌雄配子的传递规律。同时,利用"双顶交"法改良易位系的综合农艺性状。基因组原位杂交结果表明, 6RS/6AL易位染色体在传递过程中结构稳定。6RS/6AL易位染色体可以高频率地通过雌、雄配子传递,其传递率分别为45.05%~53.33%和43.94%~53.04%。初步分析"双顶交"F2分离群体表明, 6RS/6AL易位染色体对主要农艺性状如株高、穗长、小穗数和自交结实率没有明显的不利影响。用"双顶交"法可以快速地改良易位系的综合农艺性状。

Cytological and genetic analyses of a wheat-rye 2RL ditelosomic addition line with adult plant resistance to powdery mildew

Rye(Secale cereale genome RR),a close relative of common wheat,possesses valuable resistance genes for wheat improvement.Due to the co-evolution of pathogen virulence and host resistance,some resistance genes derived from rye have lost effectiveness.Development and identification of new,effective resistance genes from rye is thus required.In the current study,wheat-rye line WR56 was produced through distant hybridization,embryo rescue culture,chromosome doubling and backcrossing.WR56 was then proved to be a wheat-rye 2 RL ditelosomic addition line using GISH(genomic in situ hybridization),mc-FISH(multicolor fluorescence in situ hybridization),ND-FISH(non-denaturing FISH),mc-GISH(multicolor GISH)and rye chromosome arm-specific marker analysis.WR56 exhibited a high level of adult plant resistance to powdery mildew caused by Blumeria graminis f.sp.tritici(Bgt).This resistance was carried by the added 2 RL telosomes and presumed to be different from Pm7 which is also located on chromosome arm 2 RL but confers resistance at the seedling and adult stages.WR56 will be a promising bridging parent for transfer of the resistance to a more stable wheat breeding line.A newly developed2 RL-specific KASP(kompetitive allele specific PCR)marker should expedite that work.

秦岭黑麦75K γ-secalin亚基编码基因的克隆及序列分析

以中国春-帝国黑麦2R附加系(CS-2R)为对照,采用SDS-PAGE方法鉴定了秦岭黑麦75K γ-secalin亚基;对该亚基编码基因进行克隆测序和序列比较,结果表明:(1)秦岭黑麦75K γ-secalin(NCBI数据库登录号EU368041)具有75K γ-secalin亚基典型的氨基酸一级结构;(2)在DNA水平,秦岭黑麦同南非开普敦的栽培黑麦材料存在一个SNP位点,与欧洲的帝国黑麦存在8个SNP位点,与黑麦属其他3个种S.vavilovii、S.strictum、S.sylvestre分别存在2%、4%和6%的差异;(3)在氨基酸水平,秦岭黑麦与来自南非开普敦的栽培黑麦无差异,与欧洲的帝国黑麦差异极小,而与其他3个黑麦种的差异大,这为发掘75K γ-seca-lin基因提供了参考信息;(4)基于该基因构建的进化树能有效区分黑麦属内种间的亲缘关系,S.vavilovii种与栽培黑麦关系最近,S.sylvestre与栽培黑麦最远,而S.strictum则是介于中间的一个种,这与先前的细胞学、rDNA ITS及SSR等分析结果一致。

Isolation and characterization of an isoamylase gene from rye

Genomic DNA and cDNA sequences of an isoamylase gene were isolated and characterized from the rye genome. The full-lengths of the rye isoamylase gene are 7351 bp for genomic DNA and 2364 bp for cDNA. There are 18 exons and 17 introns in the genomic sequence, which shares a similar organization with homologous genes from Aegilops tauschii, maize, rice and Arabidopsis. Exon regions of rye and other plant isoamylase genes are more conserved than the introns. High sequence similarity(>95%) was observed in mature proteins of isoamylase genes originating from rye, Ae. tauschii, wheat and barley. The transcript profile revealed that rye isoamylase is mainly expressed in the seed endosperm with a maximum level at the middle developmental stage(15 DPA). A phylogenetic tree based on the deduced aa sequences of mature proteins from rye and other plant isoamylases indicated that rye isoamylase is more closely related to Ae. tauschii wDBE1 and wheat iso1. This is the first report on identification and characterization of the isoamylase gene from rye, making it possible to explore the roles of this enzyme for amylopectin development in rye and triticale.

Identification of genes involved in the formation of soluble dietary fiber in winter rye grain and their expression in cultivars with different viscosities of wholemeal water extract

The grain of rye(Secale cereale L.) used for baking contains a large amount of non-starch polysaccharides,making it an excellent component of functional foods. But rye grain intended for alcohol production and forage use should have a reduced content of these polysaccharides. A comprehensive parameter that can predict the best field of application for winter rye grain is the viscosity of its wholemeal water extract.However, our understanding of the genetic background underlying this key trait and associated features of rye grain is poor. By analyzing six Russian winter rye cultivars, we identified the most contrasting forms and characterized the peculiarities of their water-soluble carbohydrates capable of influencing the viscosity of water extracts. Then, using phylogenetic and transcriptomic analyses, we identified in the rye genome many genes encoding putative glycosyltransferases and glycosylhydrolases responsible for the synthesis and degradation of arabinoxylans, mixed-linkage glucans, cellulose, and some other polysaccharides. We determined the dynamics of m RNA abundance for these genes at three stages of kernel development. Comparisons of gene expression levels in two contrasting cultivars revealed specific members of multigene families that may serve as promising targets for manipulating non-starch polysaccharide content in rye grain. High-viscosity cultivars were characterized by up-regulation of many glycosyltransferases involved in the biosynthesis of arabinoxylans and other cell-wall polysaccharides,whereas low-viscosity cultivars showed up-regulation of several genes encoding polysaccharidedegrading enzymes.

Discrimination of Repetitive Sequences Polymorphism in Secale cereale by Genomic In Situ Hybridization-Banding

Genomic in situ hybridization banding （GISH-banding）, a technique slightly modified from conventional GISH, was used to probe the Chinese native rye （Secale cereale L.） DNA, and enabled us to visualize the individual rye chromosomes and create a universal reference karyotype of the S. cereale chromosome 1R to 7R. The GISH-banding approach used in the present study was able to discriminate S. cereale chromosomes or segments in the wheat （Triticum aestivum L.） background, including the Triticale, wheat-rye addition and translocation lines. Moreover, the GISH-banding pattern of S. cereale subsp. Afghanicum chromosomes was consistent with that of Chinese native rye cv. Jingzhou rye; whereas the GISH-banding pattern of Secale vavilovii was different from that of S. cereale, indicating that GISH-banding can be used to study evolutionary polymorphism in species or subspecies of Secale. In addition, the production and application of GISH-banding to the study of adenine-thymine-riched heterochromaUn is discussed.

普通小麦近缘物种黑麦1R、簇毛麦1V及鹅观草1Rk^(#1)染色体特异分子标记的筛选

为筛选普通小麦近缘物种黑麦1R、簇毛麦1V及鹅观草1Rk#1染色体特异分子标记,根据已定位于普通小麦第一部分同源群的EST序列设计104对STS引物,对中国春、鹅观草、黑麦及簇毛麦进行多态性分析。在104对STS引物中,有53对在对照普通小麦中国春与鹅观草、黑麦及簇毛麦之间存在多态性。利用普通小麦-黑麦1R^7R二体异附加系筛选出5对黑麦1R染色体特异标记,分别是CINAU 19-500、CINAU20-950、CINAU21-1500、CINAU22-310和CINAU23-2000;利用普通小麦-簇毛麦1V^7V二体异附加系筛选出5对簇毛麦1V染色体特异标记,分别是CINAU23-1700、CINAU24-1050、CINAU25-1650、CINAU26-500和CINAU27-620;利用鹅观草二体异附加系DA1Rk#1、异代换系DS1Rk#1(1A)、端体系DA1Rk#1L、易位系T1Rk#1S.W、长臂缺失系Del1Rk#1L筛选出5对鹅观草1Rk#1特异标记,分别是CINAU27-960、CINAU28-1360、CINAU29-480、CINAU30-560和CINAU31-520。研究表明,可以利用普通小麦的EST序列设计PCR引物,转化成STS标记,筛选普通小麦近缘物种黑麦、簇毛麦及鹅观草等染色体特异的分子标记,快速检测和追踪导入普通小麦背景中的黑麦1R、簇毛麦1V及鹅观草1Rk#1染色体或染色体片段,为深入研究普通小麦远缘杂种材料提供新的工具。

冬牧70黑麦+箭舌豌豆混播草地生物量、品质及种间竞争的动态研究

作者在位于华北农牧交错带的承德地区鱼儿山牧场中国农业大学科技攻关试验站,对冬牧70 黑麦和箭舌豌豆单播及混播的一年生人工草地进行了生物量动态、牧草品质及种间竞争的研究。结果表明,冬牧70 黑麦单播最佳刈割期为抽穗期,混播的最佳刈割期为黑麦乳熟早期,箭舌豌豆下部豆荚充满时期。最佳的混播组合为冬牧70黑麦25% + 箭舌豌豆75%和冬牧70 黑麦50% + 箭舌豌豆50% 。从整个混播群落看,冬牧70 黑麦生长快,植株高,其竞争力强于箭舌豌豆,在竞争中占优势。

小麦耐盐种质的筛选鉴定和耐盐基因的标记

通过对 40 0份材料的芽期、苗期鉴定 ,筛选出 11份耐盐性较强的普通小麦 (TriticumaestivumL .)、小麦和黑麦 (SecalecerealeL .)、小麦和延安赖草 (Leymuschinensis (Trin .)Tzvel.)杂交后代材料 ,其中耐盐性突出的材料有 :普通小麦品种“红蚂蚱”、“科遗 2 6”、“希望”(Hope) ;小麦与黑麦杂交后代材料 98_46、98_113、98_131;小麦与延安赖草杂交后代材料 98_16 0、98_16 1、98_16 3。耐盐性表现最突出的材料是 98_113和 98_16 0。细胞学鉴定和原位杂交及醇溶蛋白酸性聚丙烯酰胺凝胶电泳 (A_PAGE)分析和低分子量谷蛋白SDS_PAGE分析 ,证明 98_113是稳定的小麦 黑麦二体附加系 ,但具体附加的是黑麦的哪条染色体还不清楚 ;98_131是小麦 黑麦 1B/ 1R易位系。结合其他 1B/ 1R材料的耐盐表现 ,提出了黑麦 1R染色体短臂上存在耐盐基因的可能性。对 (98_16 0×BanacakaMska)F2 代分离群体苗期抗盐鉴定分析 ,表明在这一杂交组合中的耐盐性状可能由一个主效基因控制。应用SSR标记技术 ,筛选到了与 98_16 0耐盐性状连锁的SSR标记WMS6 7和WMS2 13,它们与耐盐基因的遗传距离分别为 13.9cM (centMorgan)和 31.0cM。结合小麦SSR图谱分析 ,将该主效抗性基因定位在 5BL上。

小麦EST-SSR标记的开发和染色体定位及其在追踪黑麦染色体中的应用

根据小麦盐胁迫诱导和茎秆组织相关EST序列开发了81对EST-SSR引物,其中67、46、18和61对分别在小麦、黑麦、簇毛麦和大麦基因组中稳定扩增,在不同小麦和大麦品种间具有多态性的引物分别有22和23对。利用小麦缺体-四体系共定位了43对引物的81个位点,其中A、B和D染色体组上分别有29、30和22个位点,涉及除4B、3D和6D外的18条染色体。此外30对引物在黑麦基因组中具有特异扩增,其中8对分别在黑麦1R、4R、5R和R7染色体上具有特异扩增,7对在多条黑麦染色体具有相同扩增。这些新标记可有效用于小麦及其近缘物种的遗传作图与比较遗传研究。