用荧光原位杂交检测小麦背景中的1B/1R易位

用黑麦总基因组ＤＮＡ做探针进行荧光原位杂交,检测小黑麦×普通小麦３个衍生系９３０５５３、９３０５６０和９３０６１２的黑麦染色质。荧光原位杂交显示这３个品系中都有１ＲＳ染色体臂与一小麦染色体臂发生易位,易位的断点在着丝点附近。在体细胞有丝分裂细胞周期的各时期中都可见到明显的杂交信号。１ＲＳ染色体臂在间期表现为线状杂交信号贯穿细胞核。Ｃ－分带表明３个受检易位系为１ＢＬ／１ＲＳ易位。     

小麦白粉病的新抗源普通小麦－簇毛麦易位系

小麦白粉病的新抗源普通小麦－簇毛麦易位系小麦白粉病（Ｅｒｙｓｌｐｈｉｇｒａｍｉｎｔｃｆ．ｓｐ．ｔｒｉｔｉｃｉ）是世界各产麦区的重要病害之一，在我国南北麦区都有发生，并日趋严重。近期我国小麦生产上所用品种的白粉病抗源主要来自黑麦，即小麦－黑麦１Ｂ／１Ｒ...     

春小麦壳针孢菌病害抗性鉴定初步结果

１９９２～１９９３年度，对２３５份春小麦、黑麦、小黑麦材料进行了抗壳针孢叶枯病（Ｓｅｐｔｏｒｉａｔｒｉｔｉｃｉ）和燕麦壳针孢菌小麦专化型（Ｓｅｐｔｏｒｉａａｖｅｎａｅｆ．ｓｐ．ｔｒｉｔｉｃｅａ）的抗性鉴定，结果表明，仅有为数不多的高抗品种，品种间抗性存在明显差异。     

基于改进的多色荧光原位杂交技术的染色体分析

为了提高多色荧光原位杂交(M-FISH)技术的鉴定效率,简化M-FISH操作程序,探索和建立新的M-FISH实验及观察体系,本研究以‘荆辉1号’（普通小麦辉县红-荆州黑麦双二倍体）、‘南农9918’（普通小麦-簇毛麦整臂易位系T6VS.6AL）、‘扬麦22’（普通小麦-簇毛麦整臂易位系T6VS.6DL）以及‘ST5V#4S-1’（普通小麦-簇毛麦小片段易位系）等为试验材料,以2个寡核苷酸探针[(GAA)₁₀、pAs1-1]和物种基因组DNA探针作混合探针进行M-FISH分析。结果表明在一次原位杂交中同时使用3种探针,既可以识别出小麦背景中所有的外源染色体或染色体片段,又可以根据(GAA)₁₀和pAs1-1在染色体上的杂交信号分布精确鉴定出小麦和外源染色体（片段）的具体身份。这种改进的M-FISH技术可有效用于小麦及其近缘植物染色体的鉴定,并有助于快速构建物种基于FISH的分子核型。     

一种利用染色体显微切割和微克隆获得小麦基因探针的方法

利用染色体显微切割和微克隆技术对普通小麦钢８２－１２２（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ ２ｎ＝４２）染色体长臂１Ｄ染色体进行切割和ＰＣＲ扩增，提出了高分子量麦谷蛋白（ＨＭＷ－ＧＳ１）１Ｄｘ５亚基基因片段，可为获得新基因特异性探针打下一定的基础。     

硬粒小麦染色体的FISH核型分析

采用FISH(荧光原位杂交)技术,分析硬粒小麦(Sauwne 20)染色体的FISH核型特点,为该种质在小麦新品种选育上的应用提供参考。结果表明,Sauwne 20包括14对染色体,Oligo-pTa 535-2红色探针信号主要分布在A组染色体上,而B组染色体上主要分布着明亮丰富Oligo-pSc 119.2-1绿色探针信号;根据这2种探针在Sauwne 20染色体上的分布特点,可以将其不同染色体进行准确地一一鉴别。Sauwne 20与中国春普通小麦的A组和B组染色体的FISH核型基本相似,但又有一定的差别,不同小麦材料间DNA重复序列表现出遗传多样性。     

小麦-黑麦代换系鉴定及其杂交诱导易位分析

小麦-黑麦易位系在小麦染色体工程中有重要的应用,由于黑麦是小麦的近缘种属,抗病、抗逆性强,具有许多优良性状基因,是改良小麦的重要基因库。代换系代换的是整条染色体,因此它的细胞学稳定,生活力强。因此,被鉴定出的小麦-黑麦易位系具有诸多优点,在经过田间筛选后可以直接用于生产。本研究应用C-分带和原位杂交的方法进行细胞学鉴定,鉴定出代换系4个,易位系2个,并对C-分带和原位杂交实验技术的主要影响因素也进行了探讨,这对小麦品种改良具有重要意义。     

390份小麦-黑麦种质材料主要农艺性状分析及优异材料的GISH与FISH鉴定

将小麦近缘属植物黑麦中的优良基因导入小麦可以拓宽小麦的遗传基础,丰富小麦的遗传变异。本研究调查并分析了390份小麦-黑麦种质材料。在这390份种质材料中,6个主要农艺性状值均有较大的极差,说明其遗传多样性丰富。与10份小麦主栽品种相比,90%以上的材料具有穗长和分蘖数的显著优势,60%以上的材料具有小穗数优势,约30%的材料穗粒数和千粒重显著高于主栽品种。利用基因组原位杂交(genomic in situ hybridization,GISH)和多色荧光原位杂交(multicolor fluorescent in situ hybridization,mc-FISH)技术,对8份农艺性状优良的代表性材料进行染色体组成分析,发现3份为六倍体小黑麦(AABBRR),2份为八倍体小黑麦(AABBDDRR),1份为1RS·1BL易位系,其余2份不具有可见的黑麦染色体或染色体片段。值得指出的是,3份六倍体小黑麦与2份八倍体小黑麦所含的黑麦染色体不完全相同。八倍体小黑麦中有1对来源于黑麦的小染色体,而六倍体小黑麦中没有类似小染色体;并且,不同材料中黑麦4R染色体端部的GISH杂交带有明显差异。本研究结果为这些小麦-黑麦种质材料进一步应用于小麦育种提供了依据。     

普通小麦贵紫麦1号染色体的FISH核型分析

采用FISH(荧光原位杂交)技术,分析普通小麦贵紫麦1号染色体的FISH核型特点,为贵紫麦1号在小麦新品种选育上的应用提供参考。结果表明,贵紫麦1号包括21对染色体,pAs 1红色探针信号主要分布在A组和D组染色体上,pSc 119.2-1绿色探针信号则主要分布在B组染色体上;根据贵紫麦1号染色体上这2种探针的分布特征,可以准确辨识每条染色体。贵紫麦1号与硬粒小麦(AABB)在2A、5A、2B、3B及5B染色体上表现出pSc 119.2-1信号分布差异,与节节麦(DD)在染色体1D、4D及5D上也存在信号分布差异,与中国春(AABBDD)在2A、5A、6B、1D及7D染色体上的信号也各有不同,说明DNA重复序列在不同小麦材料之间具有多态性。     

染色体原位杂交的发展及其应用

原位杂交是由放射性同位素或非放射性标记的核酸探针与细胞学制片上的染色体ＤＮＡ或组织切片上的ＲＮＡ杂交,通过放射自显影或酶促生色反应、抗原抗体反应检测具有特定序列的ＤＮＡ或ＲＮＡ,从而在细胞内进行外源染色体或特定序列定位。１原位杂交技术的发展原位杂交技...     

Identification of the spontaneous 7BS/7RL intergenomic translocation in one F1 multigeneric hybrid from the Triticeae tribe

The F1 AABBRH^ch hybrids studied here were produced by crosses between the Portuguese triticale cultivar 'Douro' (AABBRR) and the tritordeum line HT9 (AABBH^chH^ch). Fluorescent in situ hybridization performed with genomic DNA probes genomic in situ hybridization (GISH) from rye and Hordeum chilense allowed the unequivocal parental genomes discrimination in all hybrids. Among 55 plants, one presented a spontaneous wheat–rye translocation which was successfully detected after GISH. Recombinant chromosomes identification was made after reprobe with pTa71 and pSc119.2. Nine rDNA loci were detected by pTa71 and pSc119.2 identified the chromosome arms involved in the translocation, after comparing the observed hybridization patterns with those described by several authors. We identified the spontaneous wheat–rye translocation as being the 7BS/7RL. Many wheat–rye translocations have been found (e.g. 1BL.1RS and 1AL.1RS), but as far as we know, this is the first time that this translocation is reported. We considered it helpful for wheat breeding programmes as it could provide the transference of interesting agronomic characteristics from rye (e.g. leaf rust resistance) to wheat.     

基于寡核苷酸探针套painting的中国春非整倍体高清核型及应用

基于寡核苷酸探针套painting的染色体鉴定技术简单、经济和高效, 可以促进小麦品种及亲缘物种染色体识别和变异体鉴定, 提高染色体工程效率。我们前期开发了寡核苷酸探针套, 包含pAs1-1、pAs1-3、AFA-4、(GAA)₁₀和pSc119.2-1共5个探针。本研究通过一次荧光原位杂交(FISH), 对源于17个非整倍体的18份材料分析发现, 其中14个染色体组成正确, 可以清晰识别相应的缺体、四体和端体。还构建了基于寡核苷酸探针套涂染的、能准确识别3个基因组和7个部分同源群染色体的高清核型, 发现4个非整倍体发生变异, 其中从N5BT5D中鉴定出一个可能的小片段相互易位系T6AS·6AL-6DL和T6DS·6DL-6AL。进一步对7个地方品种、10个栽培品种(系)和1个人工合成小麦分析, 发现15条染色体存在多态性, 涉及6条B组(除4B)、5条A组(除1A和3A)和4条D组(1D、2D、4D和7D)染色体, 可以清晰识别我国小麦生产上广泛应用的3种易位类型(T1RS·1BL、T6VS·6AL及相互易位T1RS·7DL和T7DS·1BL), 省去了基因组原位杂交(GISH)程序。另外, 对5个亲缘物种分析发现, 该探针套可以识别栽培一粒小麦、硬粒小麦Langdon、荆州黑麦、长穗偃麦草(2n=2x=14)全部和中间偃麦草30条染色体, 并构建了这5个物种的核型。本研究结果证实该寡核苷酸探针套可以有效用于小麦及亲缘物种染色体鉴定, 高清晰的中国春非整倍体核型为小麦染色体工程提供了参考标准。     

Isolation of a new repetitive DNA sequence from Secale africanum enables targeting of Secale chromatin in wheat background

A genome specific DNA sequence that detects Secale africanum chromatin incorporated into wheat was developed in this study. Random amplified polymorphic DNA (RAPD) analysis was used to search for genome specific DNA sequences of S. africanum in lines, R111, “mianyang11” (MY11) and wheat-rye 1RS/1BL translocations R25 and R57. A high copy rye-specific DNA segment pSaD15₉₄₀ of the S. africanum genome was obtained. The sequence of pSaD15 did not show any significant homology to other reported sequences in databases and it is therefore a new repetitive sequence of Secale. PCR primers were designed for pSaD15₉₄₀, which amplify a clear 887 bp fragment in S. africanum but not in any wheat. The primers also amplified an 887 bp fragment in other accessions of rye, Chinese Spring-Imperial rye chromosome additions and a diverse range of material carrying different rye chromosomes or chromosomal segments. In situ hybridization showed that probe pSaD15₉₄₀ was specifically hybridized throughout all rye chromosomes arms except for the terminal regions. The advantage of the rye-specific probe developed herein compared to those of previous reports is that it has been shown to be widely applicable to other Secale species. The probe will be useful as a molecular marker for the introgression of S. africanum and other rye chromosome segments into the wheat genome.     

Fluorescence in situ hybridization polymorphism using two repetitive DNA clones in different cultivars of wheat

Twenty‐two wheat cultivars and a wheat line were analysed with two‐colour fluorescence in situ hybridization (FISH) using the pSc119.2 and pAs1 repetitive DNA clones to detect if polymorphism could be observed in the hybridization patterns of different wheat cultivars. The FISH hybridization pattern of ‘Chinese Spring’ was compared with wheat cultivars of different origins. Differences were observed in the hybridization patterns of chromosomes 4A, 5A, 1B, 2B, 3B, 5B, 6B, 7B, 1D, 2D, 3D and 4D. Although a low level of polymorphism exists in the FISH pattern of different wheat cultivars, it is possible to identify 17 pairs of chromosomes according to their hybridization patterns with these two probes. This study will help to predict the expected variation in the FISH pattern when analysing wheat genetic stocks of different origin. It is presumed that variation in hybridization patterns are caused by chromosome structural rearrangements and by differences in the amount and location of repetitive sequences in the cultivars analysed.     

Morphological and molecular analysis of androgenetic, selfed and backcrossed plants produced from a Hordeum vulgare x H, bulbosum hybrid

Anther culture (AC) was carried out on a fertile triploid hybrid between Hordeum vulgare L. (cultivated barley) and H. bulbosum L, (bulbous barley grass) to determine whether AC-derived regenerants differed from progeny obtained through selfing and backcrossing. Chromosome counts were carried out on all plants and DNA was extracted from them to prepare Southern blots for molecular analysis. To identify true recombinants, the blots were probed with rye repetitive sequence probes (pSc119.1 and pScl19.2). which hybridize strongly and specifically to H. bulbosum DNA. Twenty probes that detect single- or low-copy sequences were hybridized with Southern blots containing restricted DNA extracted from 25 AC-derived plants, 11 selfed and six backcrossed progeny that showed hybridizations with pScll9. Although restriction fragment length polymorphisms (RFLPs) were only observed using probes that map to four of the possible 14 chromosome arms, an introgression associated with chromosome 6HS was frequently observed among plants derived from AC. selfing and backcrossing. Plants from AC differed from selfed and backcrossed progeny in their chromosome number; unique RFLP bands that were occasionally observed may indicate chromosomal rearrangements.     

对基因组原位杂交信号释译可能出现的片面性——来自一个小麦易位系(A-3)中外源遗传物质鉴定的启示

利用基因组原位杂交(GISH)和种子醇溶蛋白电泳(A-PAGE)技术对一个可能携带有10倍体长穗偃麦草(Thinopyrum ponticum (Host) Liu & Wang)遗传物质的小麦新种质A-3进行了综合鉴定. 用普通小麦"中国春"(Triticum aesticum L. cv. Chinese Spring)基因组(ABD)DNA作探针, 拟鹅观草[Pseudoroegneria stipifolia(Czern e     

Characterization of three wheat cultivars possessing new 1BL.1RS wheat-rye translocations

The wheat-rye 1BL.1RS translocation chromosomes have been used widely around the world in commercial wheat ( Triticum aestivum L.) production because of the presence of several disease resistance genes and a yield enhancement factor on the rye ( Secale cereale L.) chromosome. However, the recent reports of the loss of complete effectiveness of the disease resistance genes on the most commonly used 1BL.1RS chromosome have highlighted the need to seek and deploy additional sources of disease resistance genes. Three new sibling wheat cultivars, 'CN12', 'CN17' and 'CN18', were developed carrying 1RS arms derived from the rye inbred line L155. Genomic in situ hybridization and C-banding analysis revealed that all the three cultivars contained the rye chromosome 1RS arm fused to the wheat 1BL wheat chromosome arm. The three cultivars displayed high yields and high resistance to local powdery mildew and stripe rust pathotypes. Fluorescence in situ hybridization analysis indicated the different structure of 1BL.1RS chromosome between 'CN18' and the other two cultivars. The present study provides a new 1RS resource for wheat improvement.     

Spontaneous wheat/rye translocations from female meiotic products of hybrids between octoploid triticale and wheat

Summary Heptaploid hybrids between octoploid triticale and wheat were backcrossed as female parents with wheat to examine the rye chromosome distribution in the resultant progenies using genomic in situ hybridization (GISH). One hundred and one backcross (BC) seeds were examined and whole rye chromosome additions and substitutions, wheat/rye centric and noncentric translocations and rye telocentric chromosomes were detected. Dicentric wheat/rye translocated chromosomes were also observed. Comparisons were made with previous results on the rye chromosome distribution from male gametes of the same cross and differences were found, where in the female derived population a deficit of plants with more than two rye chromosomes was apparent relative to the anther derived population.     

Reduced chromosome association between the short arms of 5H homologues in Hordeum vulgare L. at metaphase I

Rod or ring bivalents at metaphase I (MI) reflect whether crossing-over has occurred on one or both pairs of chromosome arms, respectively, during meiotic prophase (pachytene). In cultivated barley ( Hordeum vulgare ) the highest frequencies of rod bivalents were found for satellite chromosomes (either 5H or 6H). To distinguish between chromosome 5H and 6H associations, a recombinant line derived from an H. vulgare × H. bulbosum hybrid with a hemizygous introgression from H. bulbosum on the short arm of chromosome 6H was probed with 45S rDNA and pSc119.2 using fluorescent in situ hybridization and compared with non-recombinant H. vulgare. The frequencies of rod bivalents, caused by failure of the short arms of 5H chromosomes (5HS) to associate during meiosis in the recombinant line, were similar to the frequencies of rod bivalents involving the satellite chromosomes in H. vulgare. Hence, rod bivalent formation in H. vulgare satellite chromosomes is probably based on reduced association of 5HS, corresponding to a low frequency of genetic recombination. 5HS is, therefore, more likely to show linkage disequilibrium than the other chromosome arms.