来源于中国春×洛夫林10的DH群体的磷利用效率的鉴定

黑麦(Secale cereale L.)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-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(60kg P/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系的平均值介于双亲之间.前述五个性状的平均值和耐低磷值(-P/ P的相对籽粒产量)在含有1RS/1BL易位和无1RS/1BL易位的DH系之间没有差异.这表明在洛夫林10号中的1RS与磷的利用效率和耐低磷能力没有关系,1RS上可能没有磷高效基因.因此,有可能从洛夫林10号的后代中选出高品质、高效利用磷和耐低磷的优良品种.     

小麦矮秆圆粒突变体的鉴定与分析

小麦的子粒形态性状和株高与小麦的产量密切相关,是育种的重要选择目标性状。本研究通过对小麦品种偃展1号(YZ1)EMS突变体W98的农艺性状的调查与分析,发现W98的株高只有24 cm,而野生型YZ1的株高是73 cm。突变体株高的变异是由每个节间长度变短造成的,而非节间数目减少所致。相关分析表明矮秆与圆粒性状呈显著相关。利用高秆长粒的墨西哥品种10th12与突变体W98配制杂交组合,获得1544个F2∶3单株(株系),通过对分离群体的遗传分析,发现圆粒性状是由1对不完全显性基因控制的。赤霉素(GA)与油菜素内酯(BR)激素敏感性试验表明:野生型和突变体都对GA处理不敏感;不同浓度BR的展叶试验表明野生型对BR不敏感,而突变体W98对BR敏感。     

小麦SSR和EST-SSR引物对无芒雀麦的通用性分析

无芒雀麦(Bromus inermis Leyss.)具有营养价值高、产量大、利用季节长、耐寒耐旱、适应性强等优良品质。为了解小麦SSR和EST-SSR引物在亲缘植物无芒雀麦中的通用性,本研究选用位于普通小麦7个部分同源群的203对SSR引物和46对EST-SSR引物对无芒雀麦基因组DNA进行了扩增。结果显示:有137对(67.49%)SSR和30对(65.22%)EST-SSR引物对无芒雀麦可以有效扩增,平均扩增条带数分别为2.8和2.5,即小麦SSR和EST-SSR引物在无芒雀麦中具有较高的通用性;研究还发现,来自小麦B基因组和第5同源群染色体的引物在无芒雀麦中的有效扩增比率和平均扩增条带数最低。据此推断,小麦B基因组和第5同源群染色体可能与无芒雀麦基因组的亲缘关系较远。该研究对开发无芒雀麦基因组的特异分子标记、进行遗传多样性分析和功能基因定位等具有一定的参考价值。     

普通小麦多子房基因单体分析的染色体定位及双端体分析的染色体臂定位

本研究以普通小麦——“中国春”的单体系统和多子房小麦杂交,确定控制多子房性状的基因数目及关键染色体。通过单体分析,初步确定多子房性状分别受染色体5D和6B上的2个非互补的同效异位被动隐性基因所控制,分别用m_1和m_2表示。 继续用“中国春”双端体5DL、6BL和6BS分别与多子房进行正反交,在5DL、6BL与多子房的正反交F_1植株上均出现多子房表型。由此确定,控制多子房性状的m_1、m_2基因分别位于染色体5D和6B的短臂上(5DS、6BS)。     

普通小麦中一氧化氮相关因子(TaNOA)编码基因的克隆和分子生物学分析

一氧化氮是动植物体内重要的信号分子。本研究利用同源克隆技术从六倍体普通小麦中获得一个一氧化氮相关因子(TaNOA)编码基因的全长基因组和cDNA克隆。该基因具有13个外显子和12个内含子,与拟南芥以及水稻中同源基因结构相似。根据cDNA推导的氨基酸序列与拟南芥AtNOA1的序列一致性达60%以上,具备P-环GTPaseG4-G5-G1-G2-G3的排列特征和保守的序列。对其中2个内含子的测序分析表明在六倍体小麦中TaNOA至少有3个成员。进一步用中国春小麦缺体-四体材料将这3个TaNOA基因成员分别定位在第六同源群的6A、6B和6D染色体上,本研究中获得的成员定位于6B染色体上,因此将其命名为TaNOA-B1。原生质体表达实验表明,TaNOA-B1可能定位在线粒体中。TaNOA基因在小麦根、叶片中表达较高,在幼穗和小花中有少量表达,茎中几乎检测不到表达。TaNOA的转录本水平还因脱落酸或盐处理而上升,表明它可能参与小麦对非生物胁迫的反应。本研究为进一步克隆六倍体小麦中TaNOA的其他成员及研究该基因在小麦中的功能奠定了基础。     

小麦易位系1BL/1RS×7DL.7Ag的F2分子检测及其农艺和品质性状分析

以‘豫农982’(1BL/1RS易位)和wheatear(7DL.7Ag易位)杂交后代的900个F2群体及其F2∶3家系为实验材料,对F2群体进行1BL/1RS易位和7DL.7Ag易位类型分子检测,调查分析F2群体及F2∶3家系的农艺、产量性状(F2群体的农艺性状仅作参考,重点分析F2∶3家系的性状),并探讨1BL/1RS易位和7DL.7Ag易位对小麦品质的影响。结果表明:(1)STS标记Lr19130与SSR引物Xgwm428联合使用可作为共显性标记鉴定纯合7DL.7Ag易位与杂合7DL.7Ag易位,完善了7DL.7Ag易位的分子检测方法。(2)在农艺和产量性状方面,1BL/1RS易位可显著降低株高,有助于提高穗粒数和小穗数;7DL.7Ag易位在籽粒千粒重和产量上有显著的正向作用,但7DL.7Ag易位的穗粒数显著低于非7DL.7Ag易位且有延迟小麦成熟和增加株高的趋势;1BL/1RS和7DL.7Ag双重易位可同时提升小穗数和千粒重,但穗粒数减少。(3)在品质性状方面,1BL/1RS易位主要影响沉淀值、稳定时间、弱化度、最大拉伸阻力、延伸度等主要反映蛋白质质量的性状,使面筋质量显著降低,而对蛋白质含量、湿面筋含量和吸水率等主要反映蛋白质数量的性状影响较小;7DL.7Ag易位显著提高沉淀值和面团拉伸品质参数,对小麦粉质参数和糊化参数贡献不大。由此推测,将7DL.7Ag易位应用于小麦品种选育,有望突破产量瓶颈并可较好地提升品质。     

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     

小麦耐低磷相关性状的全基因组关联分析

通过对小麦耐低磷相关性状进行全基因组关联分析(GWAS,genome-wide association study),挖掘与小麦耐低磷性显著相关的单核苷酸多态性标记(SNP,single nucleotide polymorphism)位点及候选基因,为小麦耐低磷性状的遗传基础和分子机制研究提供理论参考。本试验以198份黄淮麦区小麦品种(系)为试验材料,设置低磷和正常磷营养液水培试验,利用小麦35K芯片对分布于小麦全基因组的11896个SNP,采用Q+K关联模型对小麦耐低磷性相关性状进行关联分析。结果表明,小麦耐低磷性状表现出广泛的表型变异,变异系数为15.65%~26.59%,多态性信息含量(PIC,polymorphic information content)为0.095~0.500。群体结构分析表明,试验所用自然群体可分为2个亚群,GWAS共检测到67个与小麦耐低磷相关性状显著关联的SNP位点(P≤0.001),这些位点分布在除3A、3B和3D以外的18条染色体上,单个SNP位点可解释5.826%~9.552%的表型变异。在这些显著位点中有4个SNP位点同时关联到了2个不同的耐低磷性状。对67个SNP位点进行发掘,筛选到7个可能与小麦耐低磷性有关的候选基因。TraesCS6A02G001000和TraesCS6A02G001100在锌指合成中有重要作用;TraesCS6A02G118100可能为低磷胁迫诱导基因;TraesCS5D02G536400、TraesCS1B02G154200和TraesCS5D02G536500与低磷胁迫相关酶类基因家族有关;TraesCS1D02G231200与植物DUF 538结构域蛋白有关,是植物胁迫相关调控蛋白候选基因。     

小麦Glu-B3位点LMW-GS基因的克隆及序列分析

以小麦特殊遗传材料———六倍体普通小麦阿勃二体、1A缺体、1B缺体和1D缺体,四倍体硬粒小麦墨西粒卡以及二倍体节节麦的总基因组DNA为模板,对D Ovidio等曾报道的硬粒小麦Glu-B3位点LMW-GS基因特异引物对P1(5-′tcctgagaagtgcatgacatg-3′)和P2(5-′gtaggcaccaactccggtgc-3′)进行了PCR扩增验证.结果表明,该引物对同样能特异扩增普通小麦Glu-B3位点LMW-GS基因.利用这对引物通过AS-PCR方法克隆得到优良小麦品种小偃6号1B染色体1个LMW-GS基因片段.该基因全长为1 089 bp,包含了完整的编码区和其上游318 bp的胚乳特异表达启动子区.该基因被命名为XY-Glu-B3-LMW2(GenBank登录号为DQ630442).XY-Glu-B3-LMW2的推测蛋白含256个氨基酸(包括N-端20个氨基酸的信号肽),其成熟蛋白有8个保守的Cys残基,均分布在C-末端区.XY-Glu-B3-LMW2是从小偃6号克隆到的第2个LMW-GS基因.     

TaDEP1基因在普通六倍体小麦及其供体种中的保守与分化

根据已知小麦正源基因TaDEP1 cDNA序列设计引物,成功克隆了小麦TaDEP1基因组序列,发现该基因包含5个外显子,4个内含子.通过比较该基因在六倍体普通小麦A、B、D基因组中的差异,筛选出可以区分A、B、D基因组的分子标记Ta956.以中国春缺体-四体系为材料,利用该标记将TaDEP1基因定位于小麦5A、5B和5...     

Chromosomal locations of the genes for histones and a histone gene-binding protein family HBP-1 in common wheat

The chromosomal locations of the genes in common wheat that encode the five histones and five members of the HBP (histone gene-binding protein)-1 family were determined by hybridizing their cloned DNAs to genomic DNAs of nullitetrasomic and telosomic lines of common wheat, Triticum aestivum cv. Chinese Spring. The H1 and H2a genes are located on different sets of homoeologous chromosomes or chromosome arms, namely, 5A, 5B and 5D, and 2AS, 2BS and 2DS, respectively. Genes for the other histones, H2b, H3 and H4, are found in high copy number and are dispersed among a large number of chromosomes. The genes for all members of the HBP-1 family are present in small copy numbers. Those for HBP-1a(1) are located on six chromosome arms, 3BL, 5AL, 5DL, 6AL, 6BS and 7DL, whereas those for each HBP-1a(c14), 1a(17), 1b(c1), and 1b(c38) are on a single set of homoeologous chromosome arms; 4AS, 4BL, 4DL; 6AS, 6BS, 6DS; 3AL, 3BL, 3DL; and 3AS, 3BS, 3DS, respectively. The genes for histones H1 and H2a, and for all members of the HBP-1 family except HBP-1a(1) are assumed to have different phylogenetic origins. The genes for histone 2a and HBP-1a(17) are located in the RFLP maps of chromosomes 2B and 6A, respectively. Gene symbols are proposed for all genes whose chromosomal locations have been determined.     

小麦ARF基因家族生物信息学分析及在干旱胁迫下的表达特性研究

植物生长素响应因子ARF(auxin response factor)参与调节了植物的向性运动、顶端优势、微观的分化、侧根和茎的形态发生等众多生理反应,在植物生长发育的整个过程都起到重要调控作用。本研究通过对小麦最新基因组数据进行分析,获得了61个ARF家族基因,命名为TaARFs,根据染色体编号排列为TaARF1~TaARF61,对61个TaARFs基因进行系统生物信息学分析后发现ARF家族基因结构较为复杂,外显子数量从1个到15个变化不等,除了4号染色体和5A和5B染色体之外,其余的染色体均有ARF家族基因分布。ARF家族基因大多包含B3 DNA结构域、ARF结构域(Auxin-resp)和Aux/IAA结构域;同源进化分析表明,小麦ARF家族基因的旁系同源基因数量明显多于大麦和二穗短柄草。通过拟南芥数据库比对获得14个高同源的根系发育相关的小麦ARF家族基因,利用二系杂交小麦京麦6号及父母本根系为试材进行干旱胁迫处理及实时荧光定量PCR(qPCR)筛选。结果表明,7个小麦ARF基因不同程度受到干旱胁迫诱导,其在旱胁迫下的表达量显著高于正常条件下的表达量,可能参与干旱胁迫应答;此外本研究还发现,ARF基因在F1杂交种中表达量显著高于双亲,表现出超亲表达模式,可能参与了根系抗旱杂种优势基因表达调控网络。     

Characterization of genetic loci conferring adult plant resistance to leaf rust and stripe rust in spring wheat.

Leaf (brown) and stripe (yellow) rusts, caused by Puccinia triticina and Puccinia striiformis, respectively, are fungal diseases of wheat (Triticum aestivum) that cause significant yield losses annually in many wheat-growing regions of the world. The objectives of our study were to characterize genetic loci associated with resistance to leaf and stripe rusts using molecular markers in a population derived from a cross between the rust-susceptible cultivar 'Avocet S' and the resistant cultivar 'Pavon76'. Using bulked segregant analysis and partial linkage mapping with AFLPs, SSRs and RFLPs, we identified 6 independent loci that contributed to slow rusting or adult plant resistance (APR) to the 2 rust diseases. Using marker information available from existing linkage maps, we have identified additional markers associated with resistance to these 2 diseases and established several linkage groups in the 'Avocet S' x 'Pavon76' population. The putative loci identified on chromosomes 1BL, 4BL, and 6AL influenced resistance to both stripe and leaf rust. The loci on chromosomes 3BS and 6BL had significant effects only on stripe rust, whereas another locus, characterized by AFLP markers, had minor effects on leaf rust only. Data derived from Interval mapping indicated that the loci identified explained 53% of the total phenotypic variation (R2) for stripe rust and 57% for leaf rust averaged across 3 sets of field data. A single chromosome recombinant line population segregating for chromosome 1B was used to map Lr46/Yr29 as a single Mendelian locus. Characterization of slow-rusting genes for leaf and stripe rust in improved wheat germplasm would enable wheat breeders to combine these additional loci with known slow-rusting loci to generate wheat cultivars with higher levels of slow-rusting resistance.     

Characterization of the calmodulin gene family in wheat: structure, chromosomal location, and evolutionary aspects

Calmodulin is a ubiquitous transducer of calcium signals in eukaryotes. In diploid plant species, several isoforms of calmodulin have been described. Here, we report on the isolation and characterization of calmodulin cDNAs corresponding to 10 genes from hexaploid (bread) wheat (Triticum aestivum). These genes encode three distinct calmodulin isoforms; one isoform is novel in that it lacks a conserved calcium binding site. Based on their nucleotide sequences, the 10 cDNAs were classified into four subfamilies. Using subfamily-specific DNA probes, calmodulin genes were identified and the chromosomal location of each subfamily was determined by Southern analysis of selected aneuploid lines. The data suggest that hexaploid wheat possesses at least 13 calmodulin-related genes. Subfamilies 1 and 2 were both localized to the short arms of homoeologous-group 3 chromosomes; subfamily 2 is located on all three homoeologous short arms (3AS, 3BS and 3DS), whereas subfamily 1 is located only on 3AS and 3BS but not on 3DS. Further analysis revealed thatAegilops tauschii, the presumed diploid donor of the D-genome of hexaploid wheat, lacks a subfamily-1 calmodulin gene homologue, whereas diploid species related to the progenitors of the A and B genomes do contain such genes. Subfamily 3 was localized to the short arm of homoeologous chromosomes 2A, 2B and 2D, and subfamily 4 was mapped to the proximal regions of 4AS, 4BL and 4DL. These findings suggest that the calmodulin genes within each subfamily in hexaploid wheat represent homoeoallelic loci. Furthermore, they also suggest that calmodulin genes diversified into subfamilies before speciation ofTriticum andAegilops diploid species.     

Genome‐wide association study for 13 agronomic traits reveals distribution of superior alleles in bread wheat from the Yellow and Huai Valley of China

Bread wheat is a leading cereal crop worldwide. Limited amount of superior allele loci restricted the progress of molecular improvement in wheat breeding. Here, we revealed new allelic variation distribution for 13 yield‐related traits in series of genome‐wide association studies (GWAS) using the wheat 90K genotyping assay, characterized in 163 bread wheat cultivars. Agronomic traits were investigated in 14 environments at three locations over 3 years. After filtering SNP data sets, GWAS using 20 689 high‐quality SNPs associated 1769 significant loci that explained, on average, ~20% of the phenotypic variation, both detected already reported loci and new promising genomic regions. Of these, repetitive and pleiotropic SNPs on chromosomes 6AS, 6AL, 6BS, 5BL and 7AS were significantly linked to thousand kernel weight, for example BS00021705_51 on 6BS and wsnp_Ex_c32624_41252144 on 6AS, with phenotypic variation explained (PVE) of ~24%, consistently identified in 12 and 13 of the 14 environments, respectively. Kernel length‐related SNPs were mainly identified on chromosomes 7BS, 6AS, 5AL and 5BL. Plant height‐related SNPs on chromosomes 4DS, 6DL, 2DS and 1BL were, respectively, identified in more than 11 environments, with averaged PVE of ~55%. Four SNPs were confirmed to be important genetic loci in two RIL populations. Based on repetivity and PVE, a total of 41 SNP loci possibly played the key role in modulating yield‐related traits of the cultivars surveyed. Distribution of superior alleles at the 41 SNP loci indicated that superior alleles were getting popular with time and modern cultivars had integrated many superior alleles, especially for peduncle length‐ and plant height‐related superior alleles. However, there were still 19 SNP loci showing less than percentages of 50% in modern cultivars, suggesting they should be paid more attention to improve yield‐related traits of cultivars in the Yellow and Huai wheat region. This study could provide useful information for dissection of yield‐related traits and valuable genetic loci for marker‐assisted selection in Chinese wheat breeding programme.     

Quantitative trait loci for grain yield and adaptation of durum wheat (Triticum durum Desf.) across a wide range of water availability

Grain yield is a major goal for the improvement of durum wheat, particularly in drought-prone areas. In this study, the genetic basis of grain yield (GY), heading date (HD), and plant height (PH) was investigated in a durum wheat population of 249 recombinant inbred lines evaluated in 16 environments (10 rainfed and 6 irrigated) characterized by a broad range of water availability and GY (from 5.6 to 58.8 q ha(-1)). Among the 16 quantitative trait loci (QTL) that affected GY, two major QTL on chromosomes 2BL and 3BS showed significant effects in 8 and 7 environments, with R2 values of 21.5 and 13.8% (mean data of all 16 environments), respectively. In both cases, extensive overlap was observed between the LOD profiles of GY and PH, but not with those for HD. QTL specific for PH were identified on chromosomes 1BS, 3AL, and 7AS. Additionally, three major QTL for HD on chromosomes 2AS, 2BL, and 7BS showed limited or no effects on GY. For both PH and GY, notable epistasis between the chromosome 2BL and 3BS QTL was detected across several environments.     

Dosage Effect of Lr3 Gene in the Triticum aestivum–Puccinia recondita Specific Interaction

Gene dosage effects of Lr3 gene were studied in the Triticum aestivum–Puccinia recondita tritici specific interaction. Deletion-duplication lines carrying different doses of Lr3 gene in chromosome 6B were obtained from a cross between standard Sinvalocho MA wheat and an isogenic translocated Sinvalocho MA 1BS/6BS-1BL/6BL line. Cytogenetic analysis and assessment of pathogen development revealed evidence for Lr3 gene dosage dependence in the expression of both resistance and avirulence. Increasing doses of chromosome 6B, from nullisomic to tetrasomic condition, increased the resistance, reaching the maximum at the tetrasomic level. The role of reciprocal translocations in the evolution of disease resistance genes in hexaploid wheat, the duplication of genes for resistance as a strategy to be used in plant breeding and the implications of gene-dosage dependence in host–parasite associations are discussed.