一个花粉辐射诱导的小麦-簇毛麦相互易位染色体系的分子细胞遗传学研究

利用⁶⁰Co-γ-射线处理小麦-簇毛麦6V单体添加系花粉，并给中国春授粉，在一个M1单株减数分裂中期Ⅰ检测到一个由2条小麦-簇毛麦易位染色体和一条完整小麦染色体构成的三价体，说明参与易位的2个小麦片段均来自同一条小麦染色体，推测两条易位染色体由相互易位产生。将其中涉及外源大片段的易位染色体称为外源大片段易位（large alien segment translocation, LAST），涉及外源小片段的称为外源小片段易位(small alien segment translocation, SAST)。对后代中两个易位染色体均纯合的植株（LAST’’+SAST’’, 2n = 44）进行顺次C-分带和GISH研究，结果表明外源大片段易位染色体为T7BS-6VS•6VL，外源小片段易位染色体为T6VS-7BS•7BL，易位断点分别位于7B染色体短臂约FL0.60处及6V染色体短臂约FL0.70处。在M2代群体中检测到7种染色体组成类型，比例为3(LAST’’+SAST’’)∶20(LAST’+SAST’)∶2(LAST’’+SAST’)∶1(LAST’+SAST’’)∶1LAST’∶2SAST’∶22(0型)，其中外源大片段和外源小片段易位染色体往往相伴出现。抗病鉴定结果显示抗白粉病基因位于外源大片段易位染色体T7BS-6VS•6VL上。对LAST’+SAST’型（2n = 43）M₂代单株花粉母细胞减数分裂的GISH研究结果显示，88.5%的后期Ｉ或末期Ｉ细胞中出现T6VS-7BS•7BL和T7BS-6VS•6VL的共分离。此外，在个别后期Ｉ细胞中观察到外源大片段易位染色体T7BS-6VS•6VL发生落后和着丝粒断裂现象，并在LAST’型单株（2n =42）的自交后代中筛选到一个通过着丝粒断裂-融合产生的外源小片段插入易位T7BL•6VS-7BS，这为利用外源大片段易位进一步创制携带抗病基因的小片段插入易位提供了新的思路。还分别获得了T7BS-6VS•6VL和T6VS-7BS•7BL的纯合易位系。     

Dosage Response of Rye Genes in a Wheat Background

Wheat (Triticum aestivum L.) breeders often utilize alien sources to supply new genetic variation to their breeding programs. However, the alien gene complexes have not always behaved as desired when placed into a wheat background. The introgressed genes of interest may be linked to undesirable genes, expressed at low levels or not at all. The short arm of rye (Secale cereale L.) chromosome one (1RS) contains many valuable genes for wheat improvement. In order to study rye gene response to varying copy number, wheat lines were constructed which contained zero, two or four doses of 1RS. The meiotic behavior of rye chromosome 1R, and wheat/rye translocation chromosomes, 1AL/1RS and 1BL/1RS was studied in the F₁ hybrids between wheat lines carrying 1R or the translocation chromosomes. The IRS arm was transmitted at a very high frequency; 98 % of the F₂ plants had at least one of the chromosomes with a IRS arm. In addition, 44 % of the F₂ plants received at least one copy of the chromosomes from each parent. Analysis of the meiotic behavior of the IRS arm suggested that few euploid wheat gametes were formed. Therefore, most of the pollen must have contained IRS. It is unknown whether the lack of euploid wheat pollen could account for the high transmission frequency of the rye chromosomes. There may have been differential survival of the embryos receiving the rye chromosome as well.     

The production of yellow-seeded Brassica napus (AACC) through crossing interspecific hybrids of B. campestris (AA) and B. carinata (BBCC) with B. napus

To transfer the genes for yellow seed coat from both genomes A and C to B. napus (AACC), the hexaploid of Brassica (AABBCC) was synthesised from reciprocal interspecific crosses between yellow-seeded B.campestris (AA) and B.carinata (BBCC). The hexaploid with 27 pairs of chromosomes was red-seeded which showed that genic interaction existed in the trigenomic plants for the colour of the seed coat. Hundreds of hybrid seeds were obtained from crosses between the red-seeded hexaploid and partial yellow or brown-seeded varieties of B. napus as pollen donor. The majority of the hybrid plants (AABCC) were self fertile with brown seeds. It appeared that the chromosomes of the B genome were excluded during the meiosis of the pentaploid and a high proportion of the genetically balanced AC gametes could be produced. The fertility of the F2 population was increased and even reached normal levels for some plants. Seventy-three plants with the yellow-seeded character were isolated from 2590 open-pollinated F2 plants, most with increased fertility. After two successive self-pollinations, 18 lines produced yellow seeds and no brown seeds segregated from these populations. The morphology of the novel yellow-seeded plants was basically towards B. napus. Esterase isoenzyme electrophoresis showed that the plants contained some of the genetic background of B. campestris, B. carinata and B. napus. Cytological analysis has shown that at least some yellow-seeded lines have the B.napus AACC genome composition with 38 chromosomes and normal meiotic pairing. This revised version was published online in July 2006 with corrections to the Cover Date.     

普通小麦-簇毛麦易位系T4VS·4VL-4AL的选育与鉴定

利用染色体C-分带和基因组原位杂交分析,从普通小麦-簇毛麦4V染色体二体异附加系（DA4V）与普通小麦农林26-离果山羊草3C染色体二体异附加系（DA3C）杂种后代中选育出小麦-簇毛麦纯合易位系T4VS·4VL-4AL。SSR和RFLP标记分析表明,该易位染色体包括4VS、4VL近着丝粒部分区段和4AL顶端区段;该易位系具有良好的细胞学稳定性,结实正常,为杀配子染色体诱发形成的补偿型易位;易位系T4VS·4VL-4AL高抗梭条花叶病,是小麦抗病育种新种质。     

三系杂交棉花粉育性对高温和低温胁迫的反应

为研究在高温和低温胁迫条件下的三系杂交棉花粉育性稳定性问题,利用棉花细胞质雄性不育系和恢复系,配制2个三系杂交棉组合,抗A×浙大强恢(记作：强恢F₁)和抗A×DES-HMF277(记作：弱恢F₁),并以保持系(抗B)为对照,分别进行温室控温试验和田间自然温度试验,分析三系杂交棉对高温和低温胁迫的反应和花粉育性转换(可育至不育)的临界温度。试验表明,一般三系杂交棉的花粉育性对胁迫温度的反应比保持系敏感,常常花粉散粉少和花粉活力较低。不同三系杂交棉组合对胁迫温度的抗性存在明显差异,强恢F₁明显高于弱恢F₁,与保持系的育性相似,可育花粉率和自交结铃率较高,不孕籽率较低。经可育花粉率(Y)与温度(T)的回归分析,花粉育性转换的临界温度符合Y = a (T–T_opt)²+b模型。强恢F₁育性转换的上限和下限温度分别为38.0℃和13.0℃,弱恢F₁为36.0℃和14.0℃,保持系为38.5℃和10.0℃。与低温胁迫比较,高温胁迫在我国大部分棉区更普遍,持续时间更长,对产量影响更大。提高三系杂交棉在胁迫气温条件下的花粉育性的稳定性是近期育种的重要目标。     

The Use of Monosomic Rye Addition Lines for Transferring Rye Chromatin into Bread Wheat

Hybrid plants with 21 pairs of wheat chromosomes and with a haploid rye genome were produced by backcrossing a primary octoploid triticale with its parental hexaploid wheat. Upon a second backcrossing or selfing, the rye chromosomes were eliminated rapidly. Added rye chromosomes, in varying numbers, affected the transmission rate of wheat chromosomes significantly. Loss of wheat chromosomes ranging from 0.06 to 0.35 per plant in different populations was observed. In these plants a remarkably high incidence of wheat/rye and rye/rye translocations occurred. Translocations were identified by using the C-banding technique. Among 837 analyzed plants 64 wheat/rye and 256 rye/rye translocations were identified. In different generations of backcrossing or selfing the frequency of wheat/rye translocations varied between 4.23 % and 14.67 %. All 14 rye chromosome arms were involved in translocations but with different frequencies. BC₁F₃ plants with homozygous wheat/rye translocations were isolated The results indicate that monosomic wheat/rye addition lines may be directly used as an effective means to transfer genetic material from rye into bread wheat.     

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%。     

A DNA Probe for Rapid Screening of Sugar Beet (Beta vulgaris L.) Carrying Extra Chromosomes from Wild Beets of the Procumbentes Section

Genes from wild species of the Procumbentes section can be transferred to sugar beet chromosomes via translocations. Since large translocations, including for example a gene for nematode resistance, generally result in meiotic disturbances, there is a need to select new diploid resistant beets from progenies of monosomic wild beet addition lines. A dispersed repetitive DNA probe, which is closely correlated with the resistance gene and hybridizes exclusively with wild beet DNA, appears to be highly superior to selection based on isozyme markers. Characteristic ‘fingerprints’ on the available monosomic addition lines reveal the existence of at last 5 different chromosomes in the wild species each housing a gene for nematode resistance. This probe can be used advantageously to identify individuals carrying the intact or fragmented wild beet chromosomes, or even various amounts of translocated chromatin. Strategies are discussed for the identification of new translocation types using straightforward squash dot or Southern hybridization techniques in combination with the wild beet DNA probe.     

萝卜-芥蓝异源四倍体的合成及GISH分析

通过萝卜(Raphanus sativus L.,2n=18,RR)与白花芥蓝(Brassica alboglabra Bailey,2n=18, CC)杂交,F1经秋水仙碱加倍合成萝卜-芥蓝异源四倍体（Raphanobrassica, 2n=36, RRCC）。经F₄~F₁₀代连续育性选择,F₁₀单株种子产量达32.3 g,每角粒数达14.9。基因组原位杂交显示F₁₀减数分裂行为类似于二倍体物种,表明该异源四倍体的细胞学行为已经稳定。育性观察表明,可育花粉足够各代生产种子,但低世代杂种出现高频瘪粒种子,胚珠败孕可能是其主要原因。该萝卜-芥蓝异源四倍体可以用作向油菜（B. napus L.,2n=38,AACC）转移萝卜基因的遗传桥梁。     

籼粳杂种花粉不育基因的定位

以IR36（indica）和热研2号（japonica，广亲和品种）为亲本，构建了包含180个单株的F₂群体及包括110个标记的分子连锁图谱。利用该F₂群体，进行了水稻花粉不育数量性状基因座(quantitative trait locus, QTL)的检测和遗传效应分析，共检测到3个花粉不育QTL，分别位于第3、5、7染色体上，此外，共检测到9个由雄配子引起的偏分离QTL，其中7个与ga-14和ga-11位点的配子败育类型相同。与花粉形态鉴定相比，偏分离的数据对检测F₁杂种花粉败育基因更为敏感。在第5、6染色体上控制偏分离的2个QTL位点，其杂合基因型出现的频率偏高。在qHPS-5位点，粳型纯合子表现出比杂合子和籼型纯合子更低的育性水平。本研究获得的分子标记将有助于聚合尽可能多的中性亲和基因以解决亚种间F₁杂种的花粉不育性问题。     

Cytological and Morphological Characterization of Pisum sativum and Pisum fulvum Tetraploids

Two colchicine-induced tetraploid lines, one belonging to Pisum sativum and the other to the wild species Pisum fulvum, were compared with their corresponding diploids and among themselves for chromosome number, meiotic behaviour and for pollen and pod fertility. For both lines, five generations were analyzed; the tetraploid level was maintained throughout the generations. In P. fulvum, aneuploids with 27, 29 and 30 chromosomes were observed, while in P. sativum aneuploids with 29 chromomes were found. Meiotic analysis evidenced a higher number of quadrivalents and a lower number of univalents in P. fulvum. This particular chromosomal behaviour may also have been responsible for determining the higher pollen and pod fertility in P. fulvum. Wild species of this genus seem to have better tolerance for ploidy variations.     

Cytological relationships among Eumelilotus species analysed using cytological observations of interspecific hybrids obtained by in vitro culture

Interspecific hybrid plants between Melilotus altissima (yellow flowers) and either M. alba (white flowers) or M. taurica (white flowers) and between M. suaveolens (yellow flowers) and M. wolgica (white flowers) which were inviable because of heavy chlorosis, were grown successfully by in vitro culture and reached the flowering stage. Pollen fertility of hybrid plants between M. altissima and M. alba was about 46% whereas hybrids between M. altissima and M. taurica and between M. suaveolens and M. wolgica showed about 25% and 18% of pollen fertility, respectively. All F₁s did not set any selfed seeds. Observing the pairing of chromosomes at metaphase I in the hybrid plants, it was revealed that the genomes of M. altissima and M. alba differ by a reciprocal translocation. This result could account for the partial sterility of pollen in the F₁ hybrids. On the other hand, hybrid plants between M. altissima and M. taurica and between M. suaveolens and M. wolgica showed normal pairing of chromosomes though pollen fertility of the hybrids was very low, suggesting that low pollen fertility was controlled by sterility gene(s). These results demonstrate that M. altissima and 7 other species of the subgenus Eumelilotus share the same chromosomal constitution. The only exception, M. alba, differs from the other 8 species by a segmental interchange. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cytological screening of rice TGMS lines

An investigation was undertaken to develop new stable thermo‐sensitive genic male sterility (TGMS) rice lines in intermated progenies (IMPs) of TGMS lines by using an anther clearing technique. The results indicate that both pre‐ and post‐meiotic genetic systems operate during anther development for the expression of sterility in TGMS lines. In all the TGMS lines, sterile anthers were small with empty pollen grains of irregular shape, except for TS 16, which showed pollen‐free anthers. This indicates that the sensitive stage of TS 16 is around stage IV (stamen and pistil primordia) of panicle development. Distinct differences were observed between sterile and fertile phases with respect to anther size, shape and colour of the pollen grains in TS 18 and TS 29. The pollen grains at the sterile phase were small and irregular in shape while in the fertile phase they were plump and larger with a yellow colour, establishing that the occurrence of sterility in TS 18 and TS 29 is post‐meiotic. Three distinct classes of pollen fertility percentage viz. <20%, 50‐70% and >90% were observed in IMPs. Anther clearing in IMPs showed distinct developmental patterns of pollen production with respect to distinct classes of pollen fertility. Less than 20% pollen fertility was observed in hybrids such as TS 15 × TS 16, TS 15 × Co 47, TS 18 × TS 16 and TS 18 × Co 47 which hold promise for developing new TGMS lines with a good plant type and acceptable quality.     

Pollination systems, hybridization barriers and meiotic chromosome behaviour in Nemesia hybrids

The ability of 13 Nemesia species (six annual and seven perennial) to sexually hybridize was investigated. Six of the perennial Nemesia species investigated were inter-fertile with one another. Two of the annual species, N. macroceras and N. strumosa, were inter-fertile. Thirty three crosses were successful and resulted in viable seeds. The analysis of meiotic chromosome behaviour in interspecific hybrids indicated that Nemesia chromosomes in different parental species were homeologous. No evidence of chromosome inversions or chromosome translocations was observed during meiosis in interspecific hybrids between the six perennial Nemesia species. In the hybrids produced between N. macroceras and N. strumosa, a quadrivalent was observed during meiotic metaphase I, indicating that these two species differ by a reciprocal translocation. A successful hybridization was made between N. anisocarpa (annual) and N. foetans (perennial), producing two triploid hybrids. In the unsuccessful crosses, pollen tubes were observed entering ovaries and ovules, suggesting that post-fertilization barriers were preventing sexual hybridization. Many of these crosses produced nonviable, shrunken, empty seeds, suggesting that endosperm breakdown and embryo abortion prevent interspecific hybridization in unsuccessful crosses. The manipulation of ploidy levels in N. fruticans and N. strumosa and tissue culture of N. strumosa × N. fruticans ovules failed to overcome post-fertilization barriers between these species.     

Isolation of mildew resistant wheat-rye translocation lines from a double substitution line

A powdery mildew resistant double disomic wheat-rye substitution line carrying rye chromosomes 1R and 2R was crossed with normal bread wheats. The F₂ generation was analysed cytologically by C-banding. Wheat-rye chromosome translocations involving both rye chromosomes 1R and 2R were frequent in F₂. Lines with translocations of 1R and 2R were harvested separately. After four generations of selfing and selection for mildew resistance and fertility, fully fertile resistant lines were selected and analysed cytologically. Lines with 1BL/1RS and 2BS/2RL translocations were identified. The resistance on chromosome 1RS could not be shown to be different from control varieties carrying the same rye segment, while the resistance on 2RL is much broader than the earlier known 2RL derived resistance in the line Transec. This revised version was published online in July 2006 with corrections to the Cover Date.     

芝麻同源四倍体的诱发与鉴定

在28度的条件下，用0．3％－0．5％的秋水仙素水溶液浸种24h，诱发产生芝麻同源四倍体的效果较好，芝麻同源四倍体与二倍体相比，茎秆粗壮，叶片，花器，种子较大，但生长发育缓慢，结实性较差，芝麻同源四倍体具有52条染色体，其育性与同源染色体形式有关，联会过程中形成的四价体（IV）和两个二价体（II＋II）数目越多，育性和结实率越高。     

普通小麦中国春-百萨偃麦草异染色体系的分子标记分析

综合利用HMW-Glu亚基、STS、SSR和RFLP等分子标记对普通小麦中国春、百萨偃麦草、中国春-百萨偃麦草双二倍体和11个中国春-百萨偃麦草异染色体系进行了分析。结果表明，14对SSR、10对STS引物和6个RFLP标记可以特异追踪百萨偃麦草染色质。C7-17及其后代株系C7-17-2等编码百萨偃麦草特异HMW-Glu亚基，添加染色体涉及与小麦第１部分同源群染色体部分同源的1J；1对STS、3对SSR和1个RFLP探针可以特异追踪二体附加系CH05中的百萨偃麦草染色体，并揭示最初根据分带核型确定的J3与小麦第2部分同源群染色体具有较高的部分同源性；2对STS、1个RFLP探针和1对SSR可以追踪CH09的外源染色体，并揭示最初确定的J7与小麦第3部分同源群染色体具有较高的部分同源性；1对STS和1个RFLP探针在CH03、CH04和CH34中具有相同的多态，3个附加系可能添加了相同染色体，最初确定的J₁、J₂和J_?与小麦第7部分同源群染色体具有较高的部分同源性；3对SSR引物可以特异追踪CH12中附加的大片段易位染色体和CH11中的小片段易位染色体，推测易位可能涉及同一条百萨偃麦草染色体。发现13个标记（5个STS、3个RFLP探针和5个SSR）可以追踪未涉及到的4J和5J等染色体。     

Discovery of 2n gametes in tetraploid oat Avena vaviloviana

Summary Sexual polyploidization via the action of 2n gametes (gametes with the sporophytic chromosome number) has been identified as the most important evolutionary mode of polyploidization among plant genera. This study was conducted to determine whether 2n gametes are present in the tetraploid level of the genus Avena (2n=4×=28) Twenty tetraploid Avena lines, representing four species and one interspecific hybrid, were screened for pollen grain size in order to differentiate between n and 2n pollen. Avena vaviloviana (Malz.) Mordv. line PI 412767 was observed to contain large pollen grains at a 1.0% frequency. Cytogenetic analyses of pollen mother cells of PI 412767 revealed cells with double the normal chromosome number (i.e., 56 chromosomes at metaphase I and anaphase I). The mode of chromosome doubling was found to be failure of cell wall formation during the last mitotic division that preceded meiosis. The resulting binucleate cells underwent normal meiotic divisions and formed pollen grains with 28 chromosomes. Based on the formation and function of 2n gametes, three models involving diploid and tetraploid oat lines are proposed to describe possible evolutionary pathways for hexaploid oats. If stable synthetic hexaploid oat lines could be developed by utilizing 2n gametes from diploid and tetraploid oat species through bilateral sexual polyploidization, the resulting hexaploids could be used in breeding programs for transferring genes from diploids and tetraploids to cultivated hexaploids.     

Molecular cytogenetic characterization of wheat-Thinopyrum ponticum translocations bearing blue-grained gene(s) induced by r-ray

Eight γ-irradiation-induced Triticum aestivum – Thinopyrum ponticum translocations conveying the blue aleurone were characterized using molecular cytogenetic approach. The size of alien chromosome segments was estimated by genomic in situ hybridization (GISH). The wheat chromosome segments involved in these translocations were clearly identified by two-color fluorescence in situ hybridization (FISH) with the probes of pAs1 and pSc119.2 (or pHvG38). Most of the detected translocations were reciprocal translocations involving wheat chromosomes 1B, 2D, 3A, 4A, 5B, 6B, 6D and 7A. This series of blue-grained wheat translocation lines would be useful in theoretical studies and wheat chromosome engineering breeding.     

携带抗白粉病基因Pm21的小麦–簇毛麦小片段易位染色体在不同小麦背景中的传递率及遗传稳定性

抗白粉病基因Pm21来自小麦近缘种簇毛麦。小麦一簇毛麦小片段顶端易位系NAU418(T1AS·1AL-6VS)和小片段中间插入易位系NAU419(T4BS·4BL-6VS-4BL)携带Pm21,高抗白粉病,是小麦抗病育种新种质。为了对其育种利用提供依据,以NAU418和NAU419为亲本分别与来源于不同生态区的郑麦9023等12个小麦品种杂交,杂种F_1再分别与来源于不同生态区的农艺亲本进行正、反回交,研究两种易位染色体在不同小麦背景中的遗传稳定性及其通过雌雄配子的传递规律。DNA分子原位杂交结果表明,在杂种F_1花粉母细胞减数分裂中期Ⅰ(Pollen Mother Cell,PMC MI),两种易位染色体分别可以与对应的小麦染色体配对形成棒状二价体。正、反交结果分析表明,NAU418中的小片段顶端易位染色体T1AS·1AL-6VS通过雌配子和雄配子的传递率分别为8.00%~50.98%和7.89%~45.07%,NAU419中的小片段中间插入易位染色体T4BS·4BL-6VS-4BL通过雌配子和雄配子的传递率分别为29.17%~52.38%和7.69%~47.06%。表明2个易位系中的易位染色体都可以通过雌、雄配子传递,但是其通过雄配子的传递率均显著低于通过雌配子的传递率。