Identification and Characterization of Phosphorus Use Efficiency in a Doubled Haploid Population of Chinese Spring×Lovrin No.10

It is reported that chromosome 1R of rye (Secale cereale L.) convey phosphorus use efficient gene (s), and 1RS/1BL translocation genotype Lovrin No.10 is P use efficient. So we hypothesized whether P efficient gene(s) locate on 1RS, and the high P efficiency of Lovrin No.10 is from 1RS? To test this hypothesis, we investigated the P use efficiency (PUE) of a doubled haploid (DH) population with 61 lines derived from anther culture of F₁ hybrid between Lovrin No.10 and phosphorus uptake inefficient genotype Chinese Spring to see whether PUE differs between DH line with and without 1RS/1BL translocation. Acidic polyacrylamide-gel electrophoresis (A-PAGE) of gliadin and genomic DNA in situ hybridization (GISH) were employed to discriminate 1RS/1BL translocation DH lines from the normal 1B DH lines. Among the 61 DH lines investigated, A-PAGE analysis showed that 34 lines contained the 1RS/1BL translocation chromosome, which was characterized by the presence of a 1RS-specific Sec-1 marker bands. Further verification with GISH proved that 33 in the 34 lines contained a pair of homozygous 1RS/1BL translocation chromosomes, only one line was a 1RS/1BL monosomic line. A field experiment was carried out on P deficient soil to investigate grain yield, biomass, numbers of spikes per plant (SPP), P uptake efficiency (PUpE), and P utilization efficiency (PUtE) of the DH lines and their parents under -P (nil P applied) and +P (60 kg P/hm² applied) at maturity. Results showed soil P deficiency decreased the values of the first four traits in Lovrin No.10, but were more severe for Chinese Spring. Lovrin No.10 had higher values of all the above tested traits at both -P and +P than Chinese Spring did, but had similar PUtE with Chinese Spring. These five traits segregated, and differed greatly among DH lines under both -P and +P conditions. Although the variations among DH lines exceeded the difference between the two parents, the average values of the DH lines were between the two parents. The average of the above five traits, and P deficiency tolerance (PDT) (measured by relative grain yield of -P/+P) were not different between the DH lines with and without 1RS/1BL translocation. This indicated that there was no association between 1RS and PUE and PDT in Lovrin No.10, and 1RS may not have P efficient gene(s). Therefore, in the offspring of Lovrin No.10, it is possible to combine high PUE and PDT with good quality without the negative effect of 1RS on flour quality.     

来源于中国春×洛夫林10的DH群体的磷利用效率的鉴定(英文)

黑麦(SecalecerealeL.)1R染色体上带有磷高效基因,含有1RS/1BL易位的洛夫林10号是一个磷高效的品种。证明P效率的基因是否位于1RS上和洛夫林10号的P高效基因是否来源于1RS具有重要意义。为了验证该假设正确与否,调查了一个DH群体的61个系的P利用效率,该群体来自于高效吸收磷的品种洛夫林10号和低效吸收磷的中国春的F1代花药培养,是否带有1RS/1BL易位和不含有1RS/1BL易位的DH系间的磷利用效率存在差异?应用酸性聚丙烯酰胺凝胶电泳(A-PAGE)和基因组原位杂交(GISH)鉴定DH群体中的1RS/1BL易位系。A-PAGE分析表明61个DH系中有34个含有1RS/1BL易位染色体,因为它们含有1RS特有的Sec-1醇溶蛋白带纹。进一步用GISH证明了34个系中33个含有一对1RS/1BL纯合易位,只有一个是1RS/1BL单体。田间试验在缺磷土壤中进行,调查各DH系以及它们的亲本在-P(不施P肥)和 P(60kgP/hm-2)条件下的籽粒产量、生物量、每株穗数、磷吸收效率和磷利用效率。结果表明,土壤缺磷降低洛夫林10号的前四个性状的值,但中国春的降低更加严重。洛夫林10号无论在 P和-P条件下所有测试性状的值都高于中国春,但磷的利用效率二者相似。在-P和 P条件下,所测定的五个性状存在分离,且在DH系之间有显著差异;虽然DH系间的变异超出双亲,但是所有DH系的平均值     

黑麦碱基因（Sec–1）表达缺失的1RS／1BL易位系的鉴定

用改良的Giemsa C－带技术、DNA原位杂交和酸性聚丙烯酰胺凝胶电泳（A-PAGE）对来源于小麦品种绵阳11与不同黑麦自交系远缘杂交获得的高代株系（BC1F7）的染色体结构和醇溶蛋白进行了研究。结果发现,在鉴定的200个株系中,有45个株系经C-带和A-PAGE检测均一致地发现它们含有一对1RS /1BL易位染色体,而一个株系843-1-1,C-带鉴定、原位杂交结果均证明它含有一对1RS/1BL易位染色体,但A-PAGE醇溶蛋白图谱却不具有黑麦1RS染色体臂的黑麦碱特征带,而表达出既不同于黑麦碱又不同于亲本绵阳11的醇溶蛋白带型。这一结果表明,利用不同的黑麦亲本资源,可以获得黑麦碱基因Sec-1表达缺失的新的1RS/1BL易位系。这种新的1RS/1BL易位系缺失了影响小麦品质的黑麦碱蛋白,因此是进一步研究1RS/1BL 易位对小麦品质影响的珍贵材料。研究指出,在利用外源基因的植物育种中,外源种供体材料的遗传多样性是值得重视的基因资源。     

普通小麦×大麦杂交后代中间材料的GISH及PAGE鉴定

利用基因组荧光原位杂交 (GISH)及种子贮藏蛋白聚丙烯酰胺凝胶电泳 (PAGE)对普通小麦×大麦杂交后代中间材料进行了鉴定分析。GISH结果表明 ,WBA984和WBA9812为二体小大麦异附加系 ,WBS0 2 15和WBS0 2 6 4为小大麦二体异代换系 ,WBT0 2 12 5和WBT0 2 183为端部易位系 ;种子贮藏蛋白PAGE分析表明 ,WBA9812和WBS0 2 6 4含有大麦特有的高分子量麦谷蛋白亚基和在γ区含有大麦特有的醇溶蛋白带型 ,WBA9812为大麦 5H附加系 ,WBS0 2 6 4为 1B/ 5H代换系 ,WBT0 2 12 5为 1BL/ 5HL端部易位系。     

Identification of new T1BL.1RS translocation lines derived from wheat (Triticum aestivum L. cultivar “Xiaoyan No. 6”) and rye hybridization

Two new T1BL.1RS translocation lines, 48112 and 89121, derived from cross between common wheat (Triticum aestivum L.) cultivar “Xiaoyan No. 6” and rye (Secale cereale L.) cultivar “German White”, were developed and identified by using of molecular markers and cytogenetical methods, GISH and FISH. PCR results of primers NOR-R1 specific for rye and Glu-B3 for 1BS detected the presence of 1RS chromatin and absence of 1BS, and primer for gene 1Bx14 in 1BL indicated the existence of chromosome arm 1BL in the two lines. GISH and FISH methods confirmed the replacement of chromosome arm 1BS with 1RS. Further stripe rust resistant test and quality analysis demonstrated that the new 1BL.1RS translocation lines were higher resistant to mixed races of P. striiformis Westend and observed considerable better quality than other popularized T1BL.1RS cultivars in China. The two lines have been used in wheat breeding for high-yield potential and rust resistance.     

小偃6号背景下黑麦遗传物质的荧光原位杂交分析

利用普通小麦(Triticum aestivum L.)“小偃6号”与黑麦(Secale cereale L.)品种“德国白粒”杂交,选育出“小偃6号”类型带有黑麦性状的种质材料。应用总基因组原位杂交(GISH)进行检测,在8份材料中探测到黑麦染色质的存在,其中附加系3个,代换系1个,易位系4个;进一步用荧光绿标记探针pSc119.2及荧光红标记探针pAs1的双色荧光原位杂交(FISH)技术,对其中部分品系的染色体组成进行分析鉴定,结果表明:易位系BC116-1是1RS/1BL小麦/黑麦易位系,BC152-1是涉及一条1B染色体的1RS/1BL易位系, 代换系BC97-2是2R(2D)二体代换系;附加系BC122-3附加了一条6R黑麦染色体,一条6B染色体的长臂缺失。同时,对连续的总基因组原位杂交和双色荧光原位杂交技术在小麦育种中的应用进行了讨论。     

Mapping QTLs for Phosphorus-Deficiency Tolerance at Wheat Seedling Stage

Soil phosphorus (P) deficiency is one of the major limiting factors to crop production throughout the world. It is an important strategy to breed varieties with improved P-deficiency tolerance for sustainable agriculture. The objective of this study was to map QTLs for P-deficiency tolerance in wheat, and develop molecular marker assisted selection in breeding wheat with improved P-deficiency tolerance. A doubled haploid (DH) population, consisting of 92 DH lines (DHLs) derived from P-deficiency tolerant wheat variety Lovrin 10 and P-deficiency sensitive variety Chinese Spring, was developed for mapping QTLs for P-deficiency tolerance. A genetic linkage map consisting of 34 linkage groups was constructed using 253 SSR markers. Shoot dry weight (SDW), tiller number (TN), shoot P uptake (SPU), and shoot P utilization efficiency (PUE) were investigated at seedling stage under P deficiency (−P) and sufficiency (+P) condition in two pot trials in 2002 and 2003, respectively. All traits segregated continuously in the population under either −P or +P condition. Significant positive correlations were found in between TN, SDW and SPU, whereas significant negative correlations were observed between PUE and SPU and between PUE and TN. Twenty and 19 QTLs were detected under −P and +P condition, respectively. The 39 QTLs were distributed on 21 chromosomal regions. There were three clusters of QTLs, which were associated with Xgwm25l (on chromosomes 4B), Xgwm271.2 (on chromosome 5A), and Xgwm121 (on chromosome 5D), respectively. Compared to the DHLs with all Chinese Spring alleles at the three loci, those with all Lovrin 10 alleles had, on average, much higher SPU, SDW and TN under −P condition in both trials, suggesting the importance of the three loci in controlling P-deficiency tolerance. It was interesting to find that two of the above three loci were closely linked with vernalization requirement genes VRN-A1 (on chromosome 5A) and VRN-D1 (on chromosome 5D). Potential implication of the linkage between P-deficiency tolerance and VRN genes was discussed.     

巨穗小麦种质中外源遗传物质的细胞遗传学和分子生物学鉴定

利用C分带、基因组原位杂交并结合分子生物学手段,对12份巨穗小麦种质材料中的外源遗传物质进行了检测.结果表明,12份材料染色体数均为42,其中5份材料均具有一对小麦-黑麦(Triticum aestivum-Secalecereal)1BL/1RS易位染色体和一对中间偃麦草(Agropyron intermedium Garten)染色体、3份材料只具有一对中间偃麦草染色体、3份材料只具一对1BL/1RS染色体、1份材料无1BL/1RS和中间偃麦草染色体.进一步细胞学分析表明,此中间偃麦草染色体代换了普通小麦(Triticum aestivum L.)中的2D染色体,因其良好的同源补偿性,表示为2Ai.同时对2Ai在巨穗小麦种质中存在的遗传学意义及小麦遗传改良中的应用进行了讨论.     

巨穗小麦种质中外源遗传物质的细胞遗传学和分子生物学鉴定

利用C分带、基因组原位杂交并结合分子生物学手段,对12份巨穗小麦种质材料中的外源遗传物质进行了检测。结果表明,12份材料染色体数均为42,其中5份材料均具有一对小麦-黑麦(Triticum aestivum-Secale cereal)1BL／1RS易位染色体和一对中间偃麦草(Agropyron intermedium Garten)染色体、3份材料只具有一对中间偃麦草染色体、3份材料只具一对1BL／1RS染色体、1份材料无1BL／1RS和中间偃麦草染色体。进一步细胞学分析表明,此中间偃麦草染色体代换了普通小麦(Triticum aestivum L.)中的2D染色体,因其良好的同源补偿性,表示为2Ai。同时对2Ai在巨穗小麦种质中存在的遗传学意义及小麦遗传改良中的应用进行了讨论。     

Development of a wheat genotype combining the recessive crossability alleles kr1kr1kr2kr2 and the 1BL.1RS translocation, for the rapid enrichment of 1RS with new allelic variation

The main objective of the present work was to develop a wheat genotype containing both the recessive crossability alleles (kr1kr1kr2kr2), allowing high crossability between 6x wheat and diploid rye, and the 1BL.1RS wheat/rye translocation chromosome. This wheat genotype could be used as a recipient partner in wheat–rye crosses for the efficient introduction of new allelic variation into 1RS in translocation wheats. After crossing the wheat cultivars ‘Mv Magdaléna’ and ‘Mv Béres’, which carry the 1BL.1RS translocation involving the 1RS chromosome arm from ‘Petkus’, with the line ‘Mv9 kr1’, 117 F₂ plants were analysed for crossability, ten of which had higher than 50% seed set with rye and thus presumably carried the kr1kr1kr2kr2 alleles. Four of the ten plants contained the 1BL.1RS translocation in the disomic condition as detected by genomic in situ hybridization (GISH). The wheat × rye F₁ hybrids produced between these lines and the rye cultivar ‘Kriszta’ were analysed in meiosis using GISH. 1BL.1RS/1R chromosome pairing was detected in 62.4% of the pollen mother cells. The use of fluorescent in situ hybridization (FISH) with the repetitive DNA probes pSc119.2, Afa family and pTa71 allowed the 1R and 1BL.1RS chromosomes to be identified. The presence of the 1RS arm from ‘Kriszta’ besides that of ‘Petkus’ was demonstrated in the F₁ hybrids using the rye SSR markers RMS13 and SCM9. In four of the 22 BC₁ progenies analysed, only ‘Kriszta’-specific bands were observed with these markers, though the presence of the 1BL.1RS translocation was detected using GISH. It can be concluded that recombination occurred between the ‘Petkus’ and ‘Kriszta’ 1RS chromosome arms in the translocated chromosome in these plants.     

几类异质1BL／1RS小麦雄性不育系诱导孤雌生殖性的研究

系统调查了4种异质（即偏凸,粘果,易变和二角山羊草细胞质）1BL/1RS小麦雄性不育系与其一系列异质同核系,同质异核系,异质异核系杂交,回交世代诱导孤雌生殖性的遗传变异规律,染色体分裂行为和对外表现特点,结果表明：1、异源细胞质与小麦1BL/1RS核型专一互作,有着消弱同源染色体配对,提高中期单价体细胞率的作用;2、特定细胞质背景下,专一核型内细胞单价体频率高低与诱导孤雌生殖性的频率直接相关;3、1BL/1RS易位染色体中易位片所存在系列差异以及1BL/1RS易位染色体外基因型背景不同,诱导孤雌生殖性的频率明显不同;4、提高或抑制不同杂交,回交世代间孤雌生殖频率,不育系母本或不同基因型父本有着同等重要的作用。选择最佳父母本组合杂交可明显提高或降低后代群体中的孤雌生殖性频率。     

用染色体工程法培育小麦新品种

染色体工程是培育小麦新品种的有效方法。利用5B染色体效应及定向选择,将黑麦抗条锈基因导入普通小麦,选育出的M8003品系,对条中22—29号等7个生理小种完全免疫,农艺性状良好。核型鉴定,2n=42,含4条随体染色体;Giemsa C-带和N-带分析,以及杂种F_1染色体配对分析,初步判断属于2BS/2RS端部易位和5AL/5RL部分易位。它不同于洛夫林10、13号等1B/1R类型的抗条锈基因,为一新的小麦-黑麦类型条锈抗源,而且是可供生产直接利用的优良品系。     

Production of a new wheat line possessing the 1BL.1RS wheat-rye translocation derived from Korean rye cultivar Paldanghomil

The 1BL.1RS translocations between wheat (Triticum aestivum L.) and rye (Secale cereale L.) are widely used in bread wheat breeding programs, but all modern wheat cultivars with the 1BL.1RS have shown genetic vulnerability due to one rye source – a German cultivar, Petkus. We have developed, a new 1BL.1RS wheat-rye translocation line from the backcross of the F₁ hybrid of wheat cv. Olmil and rye cv. Paldanghomil, both cultivars from Korea. The GISH technique was applied to identify the presence of rye chromatin in 467 BC₁F₆ lines selected from 77 BC₁F₅ lines. Only one line, Yw62–11, showed wheat-rye translocated chromosomes, with a somatic chromosome number of 2n=42. C-banding patterns revealed that the translocated chromosome was 1BL.1RS, showing prominent bands in the terminal and sub-terminal regions of the short arm as well as in the centromeric region and terminal region of the long arm. This new 1BL.1RS translocation line formed 21 bivalents like common wheat at meiotic metaphase I, thereby showing complete homology. Received: 28 February 2001 / Accepted: 17 April 2001     

The agronomic performance of wheat doubled haploid lines derived from wheat x maize crosses

Summary The agronomic performance of 9 doubled haploid (DH) lines of Chinese Spring, 6 DH lines of Hope, 14 DH lines of the single chromosome substitution line Chinese Spring (Hope 5 A) and their respective parents was analyzed under field conditions. Seventeen Chinese Spring DH lines derived from wheat x Hordeum bulbosum crosses were also included for comparison. No significant variation was detected in either population of Chinese Spring DH lines and neither DH population differed from its parent. The Hope DH lines differed significantly for tiller biomass, spikelet number per ear, ear grain weight and 50-grain weight. However, all the variation could be attributed to the poor performance of only one line. Chinese Spring (Hope 5 A) DH lines showed significant variation for ear emergence time, but this was probably due to genetic heterogeneity in the parental stock. Overall, the results suggest that most DH lines produced by the wheat x maize method resemble their wheat parent, and that the variation induced in DH production is likely to be similar to that found in DHs from wheat x Hordeum bulbosum crosses.     

威岭栽培黑麦抗白粉病特性导入小麦的研究

威岭黑麦(Weiling rye)是一个高抗白粉病(Erysiphe gramininis f．sp．tritici)的中国矮杆栽培黑麦。以Weiling rye作为白粉病抗源,高感白粉病小麦栽培品种My8443为母本,从Weiling rye与小麦My8443远缘杂交的BC_2F_6后代中鉴定出一个新的小麦-黑麦易位系No．147,以实现威岭黑麦白粉病抗性向普通栽培小麦的转移。No．147及其亲本的抗白粉病特性通过苗期和成株期优势生理小种混合接种和室内单生理小种接种鉴定,改良的染色体C-分带和基因组原位杂交技术(GISH。Ge- nomic in situ hybridization)被用于鉴定小麦和黑麦的染色质,酸性聚丙烯酰胺凝胶电泳(APAGE)被用于鉴定黑麦醇溶蛋白1RS特异条带,11个黑麦种属特异性标记SCM(Secale cereale marker)引物被用于扩增分析黑麦特异性简单重复序列(SSR)。研究结果证实No．147是一个新的高抗白粉病的1BL/1RS小麦-黑麦染色体易位系,并对其产生的细胞学机制进行了分析。论文对中国栽培黑麦抗性基因资源的利用和该易位系在小麦遗传育种改良中的利用价值进行了讨论。     

Characterization of progenies of Triticum aestivum- Psathyrostachys juncea derivatives by using genomic in-situ hybridization

Using genomic in-situ hybridization (GISH) technique, 7 translocation-addition lines, 6 transloca-tion and translocation-addition lines, 2 ditelosomic addition lines and 1 translocation line were identified from Triticum aestivum L.-Psathyrostachys juncea (Fisch. ) Nevski intergeneric hybrids, of which translocation-addition and translocation and translocation-addition lines were not found in other reports. No substitutions and disomic additions were detected in the hybrids and breakages occurred in all P. juncea chromosomes studied. Results have shown that the improved GISH technique is a rapid and economical method for use in this field.     

Characterization of progenies of Triticum aestivum- Psathyrostachys juncea derivatives by using genomic in-situ hybridization

Using genomicin-situ hybridization (GISH) technique, 7 translocation-addition lines, 6 translocation and translocation-addition lines, 2 ditelosomic addition lines and 1 translocation line were identified fromTriticum aestivum L. -Psathyrostachys juncea (Fisch.) Nevski intergeneric hybrids, of which translocation-addition and translocation and translocation-addition lines were not found in other reports. No substitutions and disornic additions were detected in the, hybrids and breakages occurred in allP. juncea chromosomes studied. Results have shown that the improved GISH technique is a rapid and economical method for use in this field.     

Characterization of progenies ofTriticum aestivum-Psathyrostachys juncea derivatives by using genomicin-situ hybridization

Using genomicin-situ hybridization (GISH) technique, 7 translocation-addition lines, 6 translocation and translocation-addition lines, 2 ditelosomic addition lines and 1 translocation line were identified fromTriticum aestivum L. -Psathyrostachys juncea (Fisch.) Nevski intergeneric hybrids, of which translocation-addition and translocation and translocation-addition lines were not found in other reports. No substitutions and disornic additions were detected in the, hybrids and breakages occurred in allP. juncea chromosomes studied. Results have shown that the improved GISH technique is a rapid and economical method for use in this field.     

青海省小麦品种中Yr10和Yr15基因及其1BL/1RS易位的分子检测

利用抗条锈病基因Yr10和Yr15的SCAR和Barc8标记以及1BL/1RS易位的复合标记,对青海省育成和引进的137份小麦品种进行检测,以明确Yr10和Yr15基因以及1BL/1RS易位在青海小麦品种资源中的分布.结果显示:在137份材料中,有4份检测到Yr10基因,19份检测到Yr15基因,分别占参试材料的2.9%和13.9%,没有检测到同时携带Yr10和Yr15基因的材料;有22份材料为1BL/1RS易位,占参试材料的16.1%.研究表明,青海省大部分小麦抗锈品种及1BL/1RS易位品种为外引种品种.     

柱穗山羊草2C染色体诱发中国春小麦背景中黑麦1R染色体结构变异的研究

当柱穗山羊草(Aegilops cylindrica Host．)2C染色体单体添加到普通小麦品种中国春和以中国春为背景的派生系时,减数分裂时,不含2C染色体的配子会发生染色体结构变异。为了制备一套黑麦1R染色体缺失系以用于定位黑麦1R染色体上的控制重要农艺性状的基因,把一条2C染色体导人到小黑麦1R二体附加系(21″ 1R″)中,然后让这些个体(21″ 1R″ 2C′,2n=45)自交,以便产生1R染色体结构变异体。实验共检测了345粒F,种子,83粒种子带有结构变异的黑麦1R染色体(24．1％)。通过C分带和原位杂交检测,对来自于23株F2的46个F3植株所带有的异常1R染色体进行了归类：其中1RL端体为39．1％,1RL等臂染色体为2．2％,1RL易位系为32．6％。1RS端体为4．3％,1RS等臂染色体为4．3％,切点在长臂上的缺失体为2．2％。在6．5％的植株中同时含有2种类型的1R染色体结构变异。其余8．7％带有异常1R染色体的个体因为没有原位杂交结果而无法判断是属于哪种类型。已获得的1R结构变异株将有可能进一步发展成为一套可用于定位黑麦1R染色体上重要功能基因的遗传材料。另外,还探讨了综合应用细胞学和分子标记方法鉴定易位染色体中小麦染色体片段的尝试,并对所获结果进行了讨论。