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。     

硬粒小麦-节节麦人工合成种HMW-GS组成及1.5+10亚基的遗传分析

对C IMM YT的99份硬粒小麦—节节麦人工合成种(简称合成种)的HMW-G S组成分析发现,G lu-B 1和G lu-D 1位点的变异类型比普通小麦丰富,分别有9种和12种亚基类型;筛选出含有比5 10亚基更优质的1.5 10和5 12亚基的合成种分别有8份和1份;含有优质亚基1.5 10的合成种与普通小麦杂交结实正常;对2个合成种与2个普通小麦品种的8个正反交组合F1种子电泳发现,优质亚基1.5 10在F1代能正常表达,双亲所有亚基在F1代都得到表达,表现共显性遗传.本研究为优质亚基1.5 10和5 12转育到普通小麦中奠定了基础.     

小麦-黑麦代换系5A/5R与6A/6R杂交诱导同祖染色体配对与易位的研究

利用两个小麦-黑麦异源双代换系DS 5A/5R与DS 6A/6R杂交,探讨同祖染色体配对的可能性与创制小麦黑麦异源易位系.在方法上对杂种F1的减数分裂行为进行研究,观察5R与5A、6R与6A配对频率,探讨同祖染色体配对规律.实验结果看到杂交F1减数分裂中有22.91%的花粉母细胞有小麦染色体(ABD组)与黑麦染色体(R组)发生同祖配对.在F2及以后世代,通过染色体C分带、原位杂交检测,选择小麦-黑麦易位系.在F2代的45株中检测到9株有易位,易位频率为20%,是目前小麦-黑麦染色体易位频率最高的.染色体易位有的来源于同祖配对的交换,有的来源于单价体错分裂或断裂的重建.     

小麦HMW-GS转基因沉默效应的遗传分析

以主栽小麦品种'高原602'和'高原142'为母本,'QQ5'为父本进行杂交,采用SDS-PAGE检测亲本、杂种F1、F2及BC1F1、BC1F1′代的HMW-GS表达情况.结果表明:'QQ5'中HMW-GS的沉默效应在杂交后代中表现为显性,在杂种F1、F2及BC1F1、BC1F1′代中遵循孟德尔遗传规律;'QQ5'自身的沉默效应对育成品种中的其他HMW-GS也有作用;在'QQ5'和杂交后代中,对HMW-GS的沉默并不影响LMW-GS的表达;在杂交F2中,出现了一些原本只在'QQ5'的野生型(bobwhite)中表达的带型,这说明'QQ5'中HMW-GS的基因组DNA并没有被破坏,其沉默机制可能在mRNA水平.     

人β_2m转基因小鼠的制备及鉴定(英文)

制备人 β2m转基因小鼠 ,研究HLA B2 70 4基因的表达 .应用显微注射将人 β2m基因注入C5 7BL 6×昆明鼠和昆明鼠×昆明鼠F1代受精卵 .出生动物及其后代经PCR筛选 ,采用斑点杂交和Southern杂交对阳性鼠基因组DNA标本进行进一步鉴定和测定整合拷贝数 ,利用RT PCR检测阳性鼠中人 β2m转基因的表达 .6只原代仔鼠及 7只它们的下一代鼠 (F1)带有人 β2m基因 .由微注射基因后移卵出生的 86只小鼠中 ,C5 7BL 6×昆明鼠杂交仔鼠 35只 ,其中 4只阳性 (11 4 % ) ,昆明鼠×昆明鼠杂交仔鼠 5 1只 ,其中 2只阳性 (3 9% ) ,含有人 β2m基因的原代鼠×昆明鼠杂交仔鼠 2 0只 ,其中 7只阳性 .整合的转基因均为单拷贝 .Southern杂交证实上述阳性鼠确有转基因整合 .阳性鼠的皮肤、结肠、睾丸和脾脏组织中均有人β2m转基因mRNA的表达 .在转基因动物制备中 ,C5 7BL 6×昆明鼠F1代明显优于昆明鼠×昆明鼠F1代 .与人HLA B2 70 4基因相比 ,人 β2m基因不易整合 ,其整合率与整合拷贝数均较低 .得到的人 β2m转基因小鼠能够将人 β2m基困传给下一代 ,并可与人HLA B2 70 4转基因鼠交配 ,研究它的致病性     

人β_2m转基因小鼠的制备及鉴定(英文)

 制备人 β2m转基因小鼠 ,研究HLA B2 70 4基因的表达 .应用显微注射将人 β2m基因注入C5 7BL 6×昆明鼠和昆明鼠×昆明鼠F1代受精卵 .出生动物及其后代经PCR筛选 ,采用斑点杂交和Southern杂交对阳性鼠基因组DNA标本进行进一步鉴定和测定整合拷贝数 ,利用RT PCR检测阳性鼠中人 β2m转基因的表达 .6只原代仔鼠及 7只它们的下一代鼠 (F1)带有人 β2m基因 .由微注射基因后移卵出生的 86只小鼠中 ,C5 7BL 6×昆明鼠杂交仔鼠 35只 ,其中 4只阳性 (11 4 % ) ,昆明鼠×昆明鼠杂交仔鼠 5 1只 ,其中 2只阳性 (3 9% ) ,含有人 β2m基因的原代鼠×昆明鼠杂交仔鼠 2 0只 ,其中 7只阳性 .整合的转基因均为单拷贝 .Southern杂交证实上述阳性鼠确有转基因整合 .阳性鼠的皮肤、结肠、睾丸和脾脏组织中均有人β2m转基因mRNA的表达 .在转基因动物制备中 ,C5 7BL 6×昆明鼠F1代明显优于昆明鼠×昆明鼠F1代 .与人HLA B2 70 4基因相比 ,人 β2m基因不易整合 ,其整合率与整合拷贝数均较低 .得到的人 β2m转基因小鼠能够将人 β2m基困传给下一代 ,并可与人HLA B2 70 4转基因鼠交配 ,研究它的致病性     

1RS-7DS.7DL小麦-黑麦小片段易位系的鉴定

黑麦(Secale cereale L., RR)是改良普通小麦(Triticum aestivum L., AABBDD)的重要基因资源,将黑麦优异基因转移到普通小麦中,是小麦品种改良的有效途经之一。文章将四川地方品种蓬安白麦子(T. aestivum L., AABBDD) 与秦岭黑麦(S. cereale cv. Qinling, RR)杂交,染色体自动加倍获得八倍体小黑麦CD-13(AABBDDRR);通过顺序FISH和GISH分析,发现该八倍体小黑麦1RS端部与7DS的端部发生相互易位,是一个携带1RS-7DS.7DL小麦-黑麦小片段易位染色体的八倍体小黑麦。利用八倍体小黑麦CD-13与四川推广小麦品种川麦42杂交、连续自交,获得包含60个株系的F₅群体;对F₅群体的58个株系进行GISH和FISH分析发现,其中13个株系含有1RS-7DS.7DL小片段易位染色体。在这13个株系中,株系811染色体数目为2n=6x=42,是稳定的1RS-7DS.7DL小片段易位系;并且1RS特异分子标记和醇溶蛋白分析表明,1RS-7DS.7DL易位染色体1RS小片段的断裂点位于分子标记IB267-IAG95之间,不包含编码黑麦碱蛋白的Sec-1位点;同时1RS-7DS.7DL小片段易位系的千粒重与川麦42相当,远远高于八倍体小黑麦CD-13,对千粒重无负作用。因此,1RS-7DS.7DL小麦-黑麦小片段易位系可作为进一步深入研究1RS小片段上的优异基因及其遗传效应的重要材料。     

鸢尾属（Iris L.）植物的杂交育种

1994－1997年在南京进行了鸢尾属（Iris L.)的种间与种内杂交育种试验,结果表明：2个种间杂交组合 亲和力极弱,F1代杂种苗生长不良,6－8周内死亡,3个种内杂杂交组合亲和力较强,F1代杂种苗生长良好,其中2个杂交组合F1代的花色由单基因控制,。未出现花色的分离;不同花色的德国鸢尾（Iris germanica L.)品种LP和PP之间的杂交组合F1代花色受多基因控制F1代杂种花色出现明显分离,从中选出了“紫金”,彩带,金舞娃,红浪,水晶球,紫云和紫盘7个新栽培品种。     

SDS-PAGE分析转基因小麦与主栽小麦杂交后代的高分子量麦谷蛋白亚基

目的:高分子量麦谷蛋白亚基(HMW-GS)1Ax1、1Dx5是对小麦面包烘烤品质有重要影响的优质亚基。将转基因小麦株系与普通小麦栽培品种常规杂交并快速筛选后代,以选育含有外源优质亚基的主栽小麦品系。方法:将分别含有1Ax1、1Dx5亚基的转基因小麦株系B102-1-2、B73-6-1与3种普通小麦主栽品种鄂恩1号、鄂麦12号、日喀则8号常规杂交,用不连续SDS-PAGE方法鉴定12组杂交组合(正反交)F1代311颗籽粒的HMW-GS。结果:不连续SDS-PAGE分析大量子代带型,能够快速鉴定筛选出具有优质亚基的株系,转基因获得的外源优质HMW-GS基因在大部分F1子代中能够共显性遗传。结论:常规杂交育种能使外源基因有效地整合进主栽小麦的基因组中,进一步分析后代遗传的稳定性和遗传规律就可以培育出优质的新品种;不连续SDS-PAGE快速筛选优质亚基的株系具有可操作性和实用性。     

甜菊不同杂交组合结实率及其F1代萌发和生长及对NaCl耐性的比较

以甜菊(Stevia rebaudiana Bertoni)的耐盐性较强品种‘中山3号’和‘守田2号’及R-A高含量品种‘中山4号’和‘守田3号’为亲本配置7个杂交组合并获得杂交种子,对种子结实率和发芽率及F1代幼苗的存活率进行统计分析,在此基础上采用砂培和水培方法比较了亲本及F1代扦插苗对NaCl胁迫的耐性.结果表明:品种间杂交组合的结实率均显著高于同系列品种间杂交及自交组合,其中‘守田2号’ב中山3号’杂交组合的结实率最高,为74.9％;7个杂交组合F1代的种子发芽率为63.8％ ～ 89.0％,差异明显;‘守田2号’ב中山4号’杂交组合F1代幼苗存活率相对较低(79.80％),其他杂交组合F1代幼苗存活率均在93％以上.砂培条件下,用100 mmol·L-1NaCl胁迫7d,各杂交组合F1代扦插苗的存活率差异不显著;随NaCl胁迫时间的延长各杂交组合F1代扦插苗的存活率均明显下降;胁迫28 d,‘守田2号’ב守田 3号’杂交组合以及‘中山3号’自交组合F1代扦插苗的存活率显著高于其他杂交组合.水培条件下,用100、150、200和250 mmol·L-1 NaCl胁迫14 d,‘守田2号’ב中山3号’、‘中山3号’ב守田2号’和‘中山3号’ב守田3号’3个杂交组合F1代扦插苗的存活率均显著高于其耐盐亲本及其他杂交组合.研究结果说明:通过杂交提高甜菊耐盐能力是可行的,而亲本的耐盐能力及亲本配置对杂交后代目标性状有较大影响;‘中山3号’ב守田2号’、‘守田2号’ב中山3号’和‘中山3号’ב守田3号’是耐盐性较强的甜菊优良杂交组合.     

远缘杂交不需幼胚培养的节节麦基因型

六倍体普通小麦(Triticum aestivum L.)是由四倍体小麦(T.turgidum L.)与二倍体节节麦(Aegilops tanschii Coss.)天然杂交然后通过染色体自然加倍形成的异源多倍体.这一起源过程是自然条件下天然发生的,它的发生需要具备一个条件:四倍体小麦与节节麦的天然杂交种子在自然条件(没有幼胚培养等)下能够正常发芽出苗.我们从22份节节麦中发现来自中东的节节麦AS60在不采用幼胚培养等人工辅助条件下,仍然很容易与四倍体小麦和普通小麦产生有生活力的杂种植株.AS60与四倍体小麦的杂交种子有50.0%(反交)及57.1%(正交)的种子,而AS60与六倍体普通小麦的杂交种子则有45.5%不需幼胚培养等措施能够正常发芽、生长.AS60的这一特征正是普通小麦起源过程需要的条件.最后探讨了这一发现对小麦遗传改良和对普通小麦起源演化研究的意义.     

Production,morphology, and cytogenetic analysis of Elymus caninus (Agropyron caninum) x Triticum aestivum F1 hybrids and backcross-1 derivatives

Summary Intergeneric hybrids were produced between common wheat, Triticum aestivum (2n=6x=42, AABBDD) and wheatgrass, Etymus caninus (Agropyron caninum) (2n=4x=28, SSHH) — the first successful report of this cross. Reciprocal crosses and genotypes differed for percent seed set, seed development and F₁ hybrid plant production. With E. caninus as the pollen parent, there was no hybrid seed set. In the reciprocal cross, seed set was 23.1–25.4% depending upon wheat genotype used. Hybrid plants were produced only by rescuing embryos 12–13 days post pollination with cv Chinese Spring as the wheat parent. Kinetin in the medium facilitated embryo germination but inhibited root development and seedling growth. The hybrids were vigorous, self sterile, and intermediate between parents. These had expected chromosome number (2n=5x=35, ABDSH), very little chromosome pairing (0.51 II, 0.04 III) and some secondary associations. The hybrids were successfully backcrossed with wheat. Chromosome number in the BC₁ derivatives varied 54–58 with 56 as the modal class. The BC₁ derivatives showed unusually high number of rod bivalents or reduced pairing of wheat homologues. These were sterile and BC₂ seed was produced using wheat pollen.     

普通小麦×东方旱麦草属间杂种的产生及无性系的建立

本研究以普通小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．;２ｎ＝６ｘ＝４２,ＡＡＢＢＤＤ）为母本,以东方旱麦草（Ｅｒｅｍｏｐｙｒｕｍｏｒｉｅｎｔａｌｅ（Ｌ．）Ｊａｕｂ．ｅｔＳｐａｃｈ;２ｎ＝４ｘ＝２８）为父本,首次成功地获得了属间远缘杂种Ｆ１,其平均结实率为０．０８％。利用植物细胞工程技术,对杂种幼胚愈伤组织的诱导、胚性无性系的建立、植株再生、壮苗培养等,最终获得了生长正常的杂种Ｆ１植株。同时,通过对杂种幼胚愈伤组织、根尖细胞的细胞学观察,结果表明该杂种为真杂种,即２ｎ＝５ｘ＝３５（预期染色体数）的杂种细胞占主体;另外,因组培过程中发生了染色体数目的变异,故也有少量２ｎ＝２８－３４染色体数的细胞。以上杂种的获得为将旱麦草优异基因向小麦的转移奠定了基础。     

Transfer of leaf rust and stem rust resistance genes from Triticum triaristatum to durum and bread wheats and their molecular cytogenetic localization.

Six accessions of Triticum triaristatum (Willd) Godr. &Gren. (syn. Aegilops triaristata) (6x, UUMMUnUn), having good resistance to both leaf rust (Puccinia recondita f.sp. tritici Rob. ex Desm) races and stem rust (P. graminis f.sp. tritici Eriks. &Henn.) races, were successfully crossed with both susceptible durum wheats (T. turgidum var. durum L., 2n = 28, AABB) and bread wheats (T. aestivum, 2n = 42, AABBDD). In some crosses, embryo rescue was necessary. The T. triaristatum resistance was expressed in all F1 hybrids. Backcrossing of the F1 hybrids to their wheat parents to produce BC1F1 plants was more difficult (seed set 0-7.14%) than to produce F1 hybrids (seed set 12.50-78.33%). The low female fertility of the F1 hybrids was due to low chromosome pairing. Only gametes with complete or nearly complete genomes from the F1 hybrids were viable. In BC2F4 populations from the cross MP/Ata2//2*MP, monosomic or disomic addition lines (2n = 21 II + 1 I or 22 II) with resistance to leaf rust race 15 (IT 1) were selected. In BC2F2 populations from the crosses CS/Ata4//2*MP and MP/Ata4//2*MP, monosomic or disomic addition lines with resistance to either leaf rust race 15 or stem rust race 15B-1 (both IT 1) were selected. Rust tests and cytology on the progeny of the disomic addition lines confirmed that the genes for rust resistance were located on the added T. triaristatum chromosomes. The homoeologous groups of the T. triaristatum chromosomes in the addition lines from the crosses MP/Ata2//2*MP, CS/Ata4//2*MP, and MP/Ata4//2*MP were determined to be 5, 2, and 7, respectively, through the detecting of RFLPs among genomes using a set of homoeologous group specific wheat cDNA probes. The addition lines with resistance to leaf rust race 15 from the crosses MP/Ata2//2*MP and CS/Ata4//2*MP were resistant to another nine races of leaf rust and the addition line with resistance to stem rust race 15B-1 from the cross MP/Ata4//2*MP was resistant to another nine races of stem rust as were their T. triaristatum parents. Since such genes provide resistance against a wide spectrum of rust races they should be very valuable in wheat breeding for rust resistance.     

Meiotic restitution in wheat polyhaploids (amphihaploids): a potent evolutionary force

Polyploidy is well recognized as a major force in plant speciation. Among the polyploids in nature, allopolyploids are preponderant and include important crop plants like bread wheat, Triticum aestivum L. (2n = 6x = 42; AABBDD genomes). Allopolyploidy must result through concomitant or sequential events that entail interspecific or intergeneric hybridization and chromosome doubling in the resultant hybrids. To gain insight into the mechanism of evolution of wheat, we extracted polyhaploids of 2 cultivars, Chinese Spring (CS) and Fukuhokomugi (Fuko), of bread wheat by crossing them with maize, Zea mays L. ssp. mays. The derived Ph1-polyhaploids (2n = 3x = 21; ABD) showed during meiosis mostly univalents, which produced first-division restitution (FDR) nuclei that in turn gave rise to unreduced (2n) male gametes with 21 chromosomes. The haploids on maturity set some viable seed. The mean number of seeds per spike was 1.45 +/- 0.161 in CS and 2.3 +/- 0.170 in Fuko. Mitotic chromosome preparations from root tips of the derived plantlets revealed 2n = 42 chromosomes, that is, twice that of the parental polyhaploid, which indicated that they arose by fusion of unreduced male and female gametes formed by the polyhaploid. The Ph1-induced univalency must have produced 2n gametes and hence bilateral sexual polyploidization and reconstitution of disomic bread wheat. These findings highlight the quantum jump by which bread wheat evolved from durum wheat in nature. Thus, bread wheat offers an excellent example of rapid evolution by allopolyploidy. In the induced polyhaploids (ABD) that are equivalent of amphihaploids, meiotic phenomena such as FDR led to regeneration of parental bread wheat, perhaps a simulation of the evolutionary steps that occurred in nature at the time of the origin of hexaploid wheat.     

普通小麦与祖山羊草杂种后代的细胞遗传学初步研究

粗山羊草(Aegilops tauschii(Coss.)Schmal.,DD,2n=14)是普通小麦(Triticumaestivum L.,AABBDD,2n=42)D染色体组的供体。粗山羊草中含有丰富的抗病,抗虫,抗逆,优质等优异基因,因此粗山羊草是改良普通小麦的宝贵遗传资源。但有关普通小麦与粗山羊草杂交的研究报道较少。本文试图通过对普通小麦与粗山羊草杂种后代的细胞遗传学和染色体分离规律的研究,探讨转移粗山羊草优异基因的方法和途径。     

Production,morphology, and cytogenetics of Triticum aestivum (L.) Thell × Elymus scabrus (R. Br.) Love intergeneric hybrids obtained by in ovulo embryo culture

Summary Intergeneric hybrids were produced between common wheat, Triticum aestivum (2n=6x=42, AABBDD), and an apomictic Triticeae species, Elymus scabrus (syn. Agropyron scabrum) (2n=6x=42, HHSSSS), the first successful report of this cross. Nine tiny, underdeveloped, and structureless embryos were obtained in vitro only by in ovulo embryo culture at 4 days after pollination, which gave rise to five mature hybrid plants. All the hybrid plants were vigorous and possessed a phenotype intermediate to the two parents. There were 2n=6x=42 (ABDHSS) somatic chromosomes in the hybrids. There was little or no homology between the parental genomes, as shown by an overall meiotic chromosome association of 32.83 I + 4.08 rod II + 0.21 ring II + 0.18 III + 0.02 IV. The hybrids were completely sterile and so far backcrosses to wheat parent have not been successful. Alternate approaches to induce gene transfer(s) from E. scabrus to wheat are being attempted.Agriculture Canada Contribution No. 398.     

New hybrids between Agropyron and wheat

Summary Intergeneric hybrids of Triticum aestivum (2n=42,AABBDD) with Agropyron ciliare (2n= 28,SSYY), A. trachycaulum (2n=28,SSHH), A. yezoense (2n=28,SSYY) and A. scirpeum (2n=28) are reported for the first time. F₁ hybrids of T. aestivum were also produced with A. intermedium (2n=42,E₁E₁E₂E₂Z₁Z₁) and A. junceum (2n=14,JuJu). All wheat-Agropyron hybrids were obtained by embryo rescue technique. Cultivars and reciprocal crosses differed for seed set, seed development and F₁ plant production. The F₁ hybrids were sterile. Attempts to obtain amphiploids were unsuccessful. However, backcross derivatives were obtained with wheat as the recurrent parent.The level of chromosome pairing in A. trachycaulum x wheat, A. yezoense x wheat and wheat x A. junceum hybrids provided no evidence of homologous or homoeologous pairing. Mean pairing frequencies in A. ciliare x wheat, wheat x A. scirpeum and wheat x A. intermedium hybrids indicated homoeologous or autosyndetic pairing. Ph gene was more effective in regulating homoeologous pairing in A. yezoense x wheat hybrids than in A. ciliare x wheat hybrid. Chromosome pairing data of BC₁ derivatives indicated that either some of the wheat chromosomes were eliminated or Agropyron chromosomes caused reduced pairing of wheat homologues.Contribution No. 82-653-J, Department of Plant Pathology, Kansas State Agricultural Experiment Station, Manhattan, Kan, USA     

Meiotic restriction in emmer wheat is controlled by one or more nuclear genes that continue to function in derived lines

Highly fertile F₁ hybrids were made between Triticum turgidum L. ssp. turgidum (2n = 28, AABB) and Aegilops tauschii Coss. (2n = 14, DD) without embryo rescue and hormone treatment. The F₁ plants had an average seed set of 25%. Approximately 96% of the F₂ seeds were able to germinate normally and about 67% of the F₂ plants were spontaneous amphidiploid (2n = 42, AABBDD). Cytological analysis of male gametogenesis of the F₁ plants showed that meiotic restitution is responsible for the high fertility. A mitosis-like meiosis led to meiotic restitution at either of the two meiotic divisions resulting in unreduced gametes. Test crosses of the T. t. turgidum–Ae. tauschii amphidiploid with Ae. variabilis and rye suggested that the mitosis-like meiosis is controlled by one or more nuclear genes that continue to function in derived lines. This discovery indicates a potential application of such genes in producing double haploids.     

Cytogenetic analysis of hybrids derived from wheat and <Emphasis Type="Italic">Tritipyrum</Emphasis> using conventional staining and genomic <Emphasis Type="Italic">in situ</Emphasis> hybridization

The new salt tolerant cereal, Tritipyrum (2n=6x=42, AABBE^bE^b) offers potential to introduce desirable characters for wheat improvements. This study was aimed to generate a segregating population from Iranian local wheat cultivars (2n=6x=42, AABBDD) and Tritipyrum crosses, study of the meiotic behaviour in F₂ hybrids and identification of E^b chromosomes in F₃ individuals. Results showed meiotic abnormalities in F₂ plants and different pairing frequency in the meiosis among F₂ plants. Genomic in situ hybridization revealed that total and E^b chromosome number of F₃ seeds ranged from 39 to 45 and 0 to 10, respectively. A significant prevalence of hyper-aneuploidy was observed among F₃ genotypes. C-banding patterns identified E^b chromosomes in Tritipyrum, indicating that it also can be useful to study wheat-Tritipyrum derivatives.