小麦-黑麦代换系间、代换系与易位系间杂交后代染色体易位系的选育

为了选育有经济价值的易位系,探讨小麦-黑麦间产生易位的频率,本研究以小麦-黑麦代换系DS5R/5A和DS6R/6A为母本,以小麦-黑麦易位系克珍(Kezhen)和带有杀配子染色体的代换系DSGC1为父本,分别进行田间杂交。结果表明:DS5R/5A、DS6R/6A与DSGC1(2S)杂交结实率低,平均为27.7%,与克珍(T3B.3R)杂交结实率高于前者,平均结实率为59.3%,二者均好于远缘杂交的结实率,这也表明带有杀配子染色体的亲本影响结实率;对选出的54株DS5R/5A×DSGC1、DS6R/6A×DSGC1杂种后代与80株DS5R/5A×克珍、DS6R/6A×克珍杂种后代进行C-分带、原位杂交和分子标记鉴定后发现,二者的易位频率分别为11.1%和8.8%,并且多为染色体端部小片段易位,这种小片段易位可能是代换系间杂交部分同源染色体交换产生的。此外,也表明杀配子染色体在引起染色体断裂后可发生染色体易位。本研究共获得T5RL.5AS、T5RS.5DL、T5RS.5BL、T6RL.6DS、6RL.6BS以及T6RS.6BL等13个易位系,平均易位频率为9.6%。     

小麦-黑麦小片段易位系新种质的分子细胞遗传学鉴定

为创制有利用价值的小麦-黑麦易位系,选育小麦遗传改良的新种质,本研究运用形态学与分子细胞遗传学结合的研究手段,对小麦-黑麦代换系6R/6A×克珍杂种后代中选育的11个品系进行鉴定。结果表明,11个品系形态表现为大穗多花、多小穗、抗倒伏、籽粒饱满度好等优良性状。其中有4个品系2-13、2-18、2-20、2-21穗长超过14 cm。品系2-18、2-21,分蘖数为7个,2-17千粒重为35.24 g,品系2-19主穗粒数为54,均超过亲本。另有8个品系结实率也优于亲本。亲本克珍为小麦-黑麦小片段易位系,带有7R和3R染色体遗传成分,分子标记和原位杂交显示,11个品系均带有R组染色体成分,分别有10个、6个和3个品系带有黑麦6R、7R和3R染色体遗传成分。有9个品系可确定为小麦-黑麦小片段易位系,易位类型为端部易位和中间易位。本研究成果对小麦育种及种质改良具有重要意义。     

小麦-黑麦代换系间杂交后代的SSR分析

为了深入了解7个大穗型小麦品系的遗传基础及更好地利用其对小麦进行遗传改良,利用SSR技术,对从小麦-黑麦代换系5R/5A与6R/6A杂交后代中选育的大穗型品系08-4、08-5、08-6、08-7、08-8、08-9、08-12进行了分析。结果表明,使用位于黑麦染色体上的17个SSR引物,在15个引物位点上,7个品系带型同普通小麦的相似。从所用引物中筛选出引物Xgwm247-6RL,在08-4、08-8、08-12和黑麦中扩增出约168bp、178bp、190bp 的三条黑麦特异片段,证明了品系08-4、08-8、08-12 中导入了黑麦6R染色体长臂的遗传物质。由于这些品系具有大穗、多小穗等优良性状,是有经济价值和利用价值的遗传材料。     

杀配子染色体在小麦-黑麦异代换系中的作用研究

利用小麦-黑麦异代换系H-13(1R/1D), H-24(5R/5A),H-33(6R/6A)与中国春-杀配子染色体二体异附加系CS-DA2C(来自山羊草Ae. cylindrica),CS-DA3C(来自山羊草Ae. triuncialis)配置杂交组合,对杀配子染色体的作用进行了研究。结果表明,杀配子染色体3C对H-13和H-24都是致死的,即有着完全的杀配子作用,但对H-33的杀配子作用则是不完全的,有着接近正常的结实率,说明不同的小麦-黑麦异代换系遗传背景对杀配子作用有很大影响,可能在品系H-33中存在对2C染色体杀配子作用的抑制基因。在以品系H-33为母本时,2个杀配子附加系F1的自交结实率差异显著,说明在相同的小麦-黑麦异代换系遗传背景下,染色体2C与3C的杀配子作用是不同的,在这2个杀配子染色体上可能带有不同的杀配子基因?这些研究结果为进一步创制小麦-黑麦易位系奠定了基础。     

利用六倍体小黑麦×普通小麦培育伴随易位的非整倍体

用核型和GiemsaC—带技术对六倍体小黑麦×普通小麦的杂种后代（F4）83C3796和83C3804混系今的非整倍体进行了分析。在74个随机抽样的细胞中，染色体数从35条到43条都有分布，比率分别是4．05％、5．40％、2．7O％、9．46％、9．46％、12．16％、12．16％、41．89％和2．70％．发生小麦染色体丢失的植株频率超过55．39％，2n－13植株出现的比率最低（占2．70％）．并从一般核型中观察到染色体游离片段和染色体“断裂一融合”过程．C一带分析观察到高频率的小麦一黑麦染色体易位，都属罗伯逊易位．易位包含了小麦的A、B、D组染色体和黑麦所有14条染色体臂，其中IRS易位频率最高（占48．3％），且全为纯合易位，其它易位染色体纯合的程度各自不同．研究中还观察到在一个细胞中有多个小麦一黑麦染色体易位共存的现象．文中还讨论了易位非整倍体在小麦育种上的利用问题。     

1B/1R易位系——“84059-4-2”的细胞学鉴定

利用 C-分带和 N-分带对普通小麦选系“84059-4-2”的根尖细胞染色体进行分析,发现其中的1B 染色体发生了变异。经带型比较,确定该变异染色体由小麦的1B 和黑麦的1R 易位而来,它包括1B 长臂及其着丝粒和1R 短臂。在中国春双端二体1B 与“84059-4-2”的杂种 F_1植株中,发现95.83%的花粉母细胞出现一个异型二价体和一个游离的短     

小麦新材料Talkrph1bRht_3和krph1bRht_3综合体的创制及其遗传分析

以中国春Talkrph1b综合体为基础材料 ,进一步将显性矮秆基因Rht3引入其遗传背景中 ,创制出两个新型小麦工具材料中国春krph1bRht3和中国春Talkrph1bRht3综合体。它们的株高 5 0～ 6 0cm ,与黑麦具有较高的杂交亲和性 ;它们与黑麦杂交获得的杂种F1 与中国春 黑麦F1 相比 ,花粉母细胞减数分裂中期I,染色体联会频率相对较高 ,单价体数明显减少 ,二价体数明显增加 ,并出现三价体和四价体 ,平均染色体交叉结数显著提高 ,表明两种新的综合体材料诱导部分同源染色体配对的能力较强。它们在利用远缘杂交和染色体工程技术向小麦转移其亲缘物种的有益基因中具有重要利用价值。     

基于寡核苷酸探针套painting的小麦“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p As1-1、p As1-3、AFA-4、(GAA)10和p Sc119.2-1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。     

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。     

‘龙麦35’与寒地多年生麦草杂交后代的分子细胞遗传分析

本研究利用黑龙江省哈尔滨地区冬季寒冷的气候特点以及形态学和分子细胞遗传学研究手段,对‘龙麦35’与6份寒地多年生麦草杂交后代F1进行选育和分析。结果表明,寒地多年生麦草同中间偃麦草染色体组构成一致,具有出色的抗寒性;寒地多年生麦草1-1-4和7-31花期同‘龙麦35’同步,杂交结实率明显高于其他组合。杂种F1为两亲中间型,植株成活率低,减数分裂时期寒地多年生麦草的染色体可同‘龙麦35’染色体发生配对,但染色体组配对不完全,单价体数量多于二价体,导致植株不育。F₁植株虽不具有抗寒性,但具有割后再生性。8个F₁株系,仅株系F₁18731结实,GISH检测结果表明,F₂18731染色体组带有27条‘龙麦35’染色体,14条麦草染色体和1条小麦-麦草易位染色体,染色体数为42条。本研究可为寒地多年生麦草染色体组的研究和利用提供理论基础。     

Chromosomal location of factors affecting content and composition of non-starch polysaccharides in wheat-rye addition lines

Summary Content of dietary fibre (DF) as well as content and composition of non-starch polysaccharides (NSP) were evaluated in disomic wheat-rye addition lines and octoploid triticale compared to their parental species — wheat (Grana) and rye (Dakowskie Zote). Large variation in the contents of NSP and DF were observed in the wheat-rye addition lines, especially in the soluble fractions. The double addition of rye chromosomes led to important transgression effects on content and composition of DF in wheat grain. Chromosomes 2R and 5R increased SDF content above the rye level, whereas the 3RS arm decreased SDF content below the wheat level. The octoploid triticale demonstrated the highest content of total arabinoxylan (AX), exceeding that of rye, while the double addition of chromosomes 4R, 6R and 6RL had an impact on high expression, comparable to that of rye content of total AX in wheat grain. Chromosomes 2R and 5R notably increased the proportion of soluble non-cellulosic glucose in the NSP fraction, in contrast to the rest of wheat-rye addition lines, octoploid triticale, wheat and rye, where AX was found to be predominant among NSP constituents. The effects of single chromosome pairs on content and composition of NSP proved to be higher than the effect of the whole rye genome in octoploid triticale. The most remarkable effect, especially considering the direction of changes, was of a 3R chromosome short arm addition. The obtained data might be of interest in breeding rye or triticale with lower viscous AX content as well as rich in soluble non-starch glucose polymers, regarded as a corrective factor in modern diseases.Abbreviations AX arabinoxylan - CV coefficient of variation - DF dietary fibre - IDF insoluble dietary fibre - NSP non-starch polysaccharides - SDF soluble dietary fibre - TDF total dietary fibre - TFA trifluoroacetic acid     

Cytological identification of 1B/1R wheat-rye translocations in winter wheat breeding lines

Summary The incorporation of rye (S. cereale L.) chromatin into winter wheat (T. aestivum L.) cultivars is often achieved via hybridization of unadapted wheat-rye translocation lines with adapted wheat germplasm. Identification of progenies possessing the translocated chromosome has traditionally involved phenotypic screening for the desired rye characteristics. In this study, the Giemsa N-banding technique was evaluated as a potential screening tool for detection of 1B/1R wheat-rye translocations. Five breeding lines were examined from the pedigree Aurora/2^*TAM W-101. The differential banding patterns of chromosome 1B contributed by TAM W-101 and chromosome 1B/1R contributed by Aurora allowed unequivocal identification of translocation genotypes. Three of the lines were found to be heterogeneous, whereby plants were homozygous for either the normal 1B or the translocated 1B/1R chromosome. The remaining two lines were observed to be homozygous and homogeneous for the translocated 1B/1R chromosome. The implication of N-banding chromosomal analyses to wheat breeding is presented.Contribution No. J-5172, Department of Agronomy, Oklahoma Agriculture Experiment Station, Oklahoma State University, Stillwater, OK74078.     

Allocation of a powdery mildew resistance locus to the chromosome arm 6RL of <Emphasis Type="Italic">Secale cereale</Emphasis> L. cv. ‘Jingzhouheimai’

Chinese cultivar Jingzhouheimai of Secale cereale L. confers resistance to several important wheat diseases, making it a useful resource for wheat improvement. Octoploid amphiploid (2n = 8x = 56; AABBDDRR) was synthesized by hybridization of Jingzhouheimai with hexaploid wheat landrace Huixianhong and doubling of the F1 chromosome number. This amphiploid was backcrossed to the wheat parent to produce wheat-rye chromosome addition lines. The existing six disomic addition lines (all except 6R) were screened with 16 rye-specific DNA markers and three putative markers for 6R were identified. These markers were used in selection of chromosome 6R addition lines, confirmed by the genomic in situ hybridization (GISH). In this manner, seven monosomic 6R and five disomic addition were selected, as well as one mono-telosomic 6RS addition, one mono-telosomic and one ditelosomic 6RL additions. In turn these 6R addition lines were used to develop additional 15 6R-specific markers of which six were allocated to the arm 6RS and nine to 6RL. Screening for the powdery mildew resistance identified chromosome arm 6RL as the carrier of the resistance locus. Therefore, DNA markers identified as specific to the 6RL arm can be used to monitor the introgression of the resistance locus into wheat.     

利用1RS特异标记和染色体原位杂交技术鉴定小麦1BL·1RS易位系

以小麦-黑麦1BL·1RS易位系(Kavkaz、山农030-1)、1AL·1RS易位系(Amigo)、荆州黑麦、八倍体小黑麦劲松49、1R-7R二体异附加系以及普通小麦中国春、辉县红、铭贤169、Chancellor等为材料,对65个黑麦1RS特异标记进行鉴定,从中筛选出8个稳定的标记,即NOR-1、SECA2/SECA3、SCSS30.2、Sec1Gene、Sec1Pro、ω-Sec-P1/P2、ω-Sec-P3/P4和IB-267,可用于检测1AL·1RS易位系或1BL·1RS易位系;另外3个特异标记O-SEC5′-A/O-SEC3′-R、IAG95-1和SCM-9可用于区别1RS来源不同的1AL·1RS和1BL·1RS易位系。利用这11个标记和染色体原位杂交技术对40份山东省近年育成小麦品种(系)进行检测,发现潍麦8号、鲁麦14、济宁13、山农664、山农优麦3号和烟农25为1BL·1RS易位系,而且是1RS的整臂易位系,未检测到1AL·1RS易位系和其他易位类型。     

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。     

普通小麦辉县红-荆州黑麦异染色体系的选育及其梭条花叶病抗性鉴定

为发掘和利用荆州黑麦所携抗梭条花叶病基因，综合利用分子细胞遗传学与分子标记技术结合多年抗性鉴定，从高感梭条花叶病小麦地方品种辉县红与荆州黑麦杂交后代（F₇~F₉）中选育出二体异附加系5个(分别添加1R、2R、R3、5R和R7)、5RS端二体异附加系1个和多重异附加代换系2个（染色体组成分别为20’’+2R(2D)’’+4R’’和19’’+1R（1B）’’+2R（2B）’’+4R’’）。鉴定表明，双二倍体荆辉1号高抗梭条花叶病，表明黑麦抗性基因可在小麦背景中稳定表达，2R、R7二体异附加系及2个含2R的多重异附加代换系均表现高抗，推测2R和R7上可能携带抗病基因。这些材料是研究荆州黑麦抗性基因遗传及小麦抗病育种的新种质。     

Development and identification of wheat–Haynaldia villosa T6DL.6VS chromosome translocation lines conferring resistance to powdery mildew

Three lines conferring resistance to powdery mildew, Pm97033, Pm97034 and Pm97035, were developed from the cross of Triticum durum-Haynaldia villosa amphidiploid TH3 and wheat cv.'Wan7107' via backcrosses, immature embryo and anther culture. Genomic in situ hybridization analysis showed that these lines were disomic translocation lines. Cytogenetic analysis indicated that the F1 plants of crosses between the three translocation lines and 'Wan7107' and crosses between the three translocation lines and substitution line 6V(6D) formed 21 bivalents at meiotic metaphase I. Aneuploid analysis with 'Chinese Spring' double ditelocentric stocks indicated that the translocated chromosomes were related to chromosome 6D. Biochemical and restriction fragment-length polymorphism (RFLP) analyses showed that the translocation lines lacked a specific band of 6VL of H. villosa compared with the substitution and addition lines but possessed specific markers on the short arm of the 6V chromosome of H. villosa. The three translocation lines lacked specific biochemical loci and RFLP markers located on chromosome 6DS. The results confirmed that Pm97033, Pm97034 and Pm97035 were T6DL.6VS translocation lines.     

Chromosomal location of aluminium tolerance genes in rye

Rye is known for its high tolerance of aluminium in the soils in comparison with wheat and other cereals. To localize the major gene/ genes controlling aluminium tolerance on the rye chromosomes, four series of wheat‐rye addition lines, two sets of triticale D(R) substitution lines and several wheat/rye translocation lines were tested in experiments on seedlings grown in nutrient solutions with various concentrations of aluminium. The results indicate that the major locus responsible for Al tolerance in rye is located on the short arm of chromosome 3R. The importance of these results for controlled introgressions into cereals is discussed.