人工合成冬性六倍体小麦的研究

以冬性四倍体硬粒小麦(Triticum durum,2n=28,AABB)为母本与粗山羊草(Aegilops tauschii,2n=14,DD)杂交,得到的单倍体幼胚(n=21,ABD)经组织培养拯救,获得的幼苗经染色体加倍而成为合成小麦(AABBDD)。从中鉴定、筛选出冬性的合成小麦。幼胚仅在1/2 MS培养基上培养,成苗率为75.81%;根据幼胚的发育状态,将发育较完善的幼胚直接接种在1/2 MS培养基上,将发育不良的幼胚先接种于1/2 MS+2 mg/L 2,4-D培养基上进一步养育幼胚,之后视幼胚发育状况再将其转入1/2 MS培养基中培养成苗,此方法的成苗率为92.44%,较前者的成苗率提高了16.63%。染色体加倍在冬季塑膜拱棚内用0.05%秋水仙素进行半根法处理,较容易获得健壮苗,并且分蘖多。     

人工合成六倍体小麦醇溶蛋白遗传多样性分析

运用酸性聚丙烯酰胺凝胶电泳(A-PAGE)技术,对96份人工合成六倍体小麦的醇溶蛋白多样性进行了分析。结果显示,96份人工合成小麦中,共分离出65条不同的醇溶蛋白谱带,其中ω区22条,β和γ区各17条,α区9条,但各醇溶蛋白在电泳图谱中出现的频率差异较大,其变化范围为1.04%~91.67%。醇溶蛋白遗传多样性指数(H′)及多态性信息含量(PIC)分析结果显示,β、ω两个谱带区醇溶蛋白组成最为丰富,而α区最低;聚类分析结果显示,材料间的平均遗传距离为0.86,在遗传距离为0.83水平上,96份材料被划分为4个主要类群,类群间的关系基本反映了合成双二倍体的亲缘关系。研究结果表明,人工合成六倍体小麦醇溶蛋白基因位点表现出广泛的遗传变异,具有丰富的遗传多样性。     

合成六倍体小麦的遗传育种

普通小麦是由四倍体小麦栽培类型与野生二倍体节节麦远缘杂交形成的异源六倍体.普通小麦保持了四倍体小麦的高产潜力,D基因组的加入丰富了食品加工产品类型、增强了环境适应能力.与二倍体作物不同,普通小麦有3个亚基因组,存在大量重复基因,基因组缓冲性、可塑性强,单个基因拷贝可能对育种改良的效果有限.小麦3个亚基因组的遗传多样性是...     

Transcriptome Sequencing of a Novel Albino Mutant of Hexaploid Sweetpotato

Plant Molecular Biology Reporter - Sweetpotato (Ipomoea batatas L), the herbaceous plant cultivated for its starchy storage roots, has a complex genome (2n?=?6x?=?90) with...     

Ribosomal RNA Cistron Multiplicity and Nucleolar Organizers in Hexaploid Wheat

The ribosomal DNA (DNA complementary to ribosomal RNA) content of twelve different wheat genotypes has been studied. Some of these genotypes are aneuploids with deletions or additions of chromosomes bearing nucleolar organisers. The rDNA contents of these genotypes provide several examples of a clear departure from a correlation between the number of rRNA cistrons and the number of nucleolar organisers. Thus the number of rRNA cistrons per nucleolar organiser is not constant in wheat. Wheat DNA was found to have a mean buoyant density of approximately 1.702 g/cc for all genotypes studied and rRNA hybridized selectively to DNA of buoyant density approximately 1.710 g/cc. The thermal stabilities of all the rRNA/DNA hybrids were essentially identical.     

Differential rRNA Genes Expression in Hexaploid Wheat Related to NOR Methylation

Ribosomal RNA genes expression was analysed in 18 Portuguese bread wheat accessions (Triticum aestivum L. em Thell.), and the number of argyrophilic-nucleolar organiser regions (Ag-NORs) per cultivar was scored. Ten accessions presented six Ag-NORs per metaphase and six nucleoli per interphase, and eight accessions presented four Ag-NORs per metaphase and four nucleoli per interphase. Fluorescent in situ hybridisation with the 45S rDNA sequence pTa71 and genomic DNA from Aegilops tauschii (2n = 2x= 14, DD) confirmed the Ag-NOR location and identified the six satellited chromosomes as being the chromosome pairs 1B, 6B and 5D. The methylation pattern of the NOR region was studied by Southern blot using pTa71 as probe, which represented a complete rDNA unit of bread wheat. DNA digestions performed by MspI and HpaII resulted in different patterns revealing the high level of cytosine methylation at their recognition sequences. The total percentage values of NOR methylation indicated that wheat accessions with a maximum of four Ag-NORs were more heavily methylated at the NOR region than accessions with a maximum of six Ag-NORs.     

K-Box Sequence of the Apetala1-Class MADS-Box Genes in Hexaploid Wheat

Direct amplification of genomic DNA from four wheat species produced DNA fragments corresponding to the K-box sequence of the apetala1/squamosa class of the MADS-box genes. Exons 3 to 5 were highly conserved within the tribe Triticeae and very similar to the apetala1 genes of darnel ryegrass, rice, and maize. Most of the variations observed were due to synonymous substitutions: the deduced amino acid sequences were 89–99% similar within the Triticeae and 88–94% within the entire family Poaceae. Introns 3 and 4 of the apetala1 class genes were similar in wheat and rye and differed from those in other MADS-box genes presently known.     

Comparative Analysis of Physio-Biochemical Responses to Cold Stress in Tetraploid and Hexaploid Wheat

The cellular changes induced by cold stress (CS) include responses that lead to oxidative stress and limits plant growth, metabolism, and productivity. In this study, responses of physio-biochemical to CS phases were comparatively studied in three genotypes of bread and durum wheats differing in sensitivity, two of them (Norstar, bread wheat and Gerdish, durum wheat) were tolerant to CS and the other one, SRN (durum wheat) was sensitive to CS. 14-day-old seedlings were subjected to CS (12 and 24 h) with or without cold acclimation (CA) phase. During CS, the elevated levels of electrolyte leakage index, contents of hydrogen peroxide (H₂O₂), and malondialdehyde in Norstar and Gerdish were lower than that of SRN plants. Positive correlation and co-regulation of reactive oxygen species (ROS) scavenging systems, superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase, and proline especially after CA phase suggested crucial role for holding back toxic ROS levels in CS phase. However, different activities of this system resulted in different intensities of oxidative stress in acclimated and non-acclimated plants. Our results showed that a CA phase induced oxidative stress tolerance by modulating antioxidative systems. These responses confirmed the existence of wide range of genetic capacity in durum wheat to increase cold tolerance particularly in Gerdish so that the sensitivity of SRN plants to CS was linearly correlated with the more decrease of antioxidant systems. These general responses may be a sign for associating other metabolites or enzymes activities to create partly tolerance against cold-induced oxidative stress. Eventually, assessing the dynamic of cell responses in short intervals after CS without CA phases profitably could be a novel path in plant stress response investigations in a short period of time.     

Comparison of Hexaploid and Tetraploid Wheat Cultivars in their Responses to Water Stress

We studied the effect of water stress imposed at anthesis and pre-anthesis stages on oxidative stress and antioxidant activity in four wheat cultivars, two hexaploid Triticum aestivum cultivars, drought resistant cv. C 306 and drought susceptible cv. Hira, and two tetraploid cultivars, T. durum cv. A 9-30-1 and T. dicoccum cv. HW 24. Water stress decreased relative water content (RWC), membrane stability index (MSI), and increased H₂O₂ and malondialdehyde (MDA) contents as well as activity of superoxide dismutase (SOD), catalase (Cat) and peroxidase (POX) in all the genotypes at all the stages. Both the tetraploid cultivars showed higher RWC, MSI and SOD activity, and lower H₂O₂ and MDA contents under water stress than hexaploid ones. Cat and POX activities were highest in C 306.     

小麦A/B染色体组SSR标记在新小麦合成前后的比较研究

微卫星分子标记已广泛用于普通小麦遗传和进化研究。由于人工合成小麦与小麦品种之间存在高的遗传多样性,人工合成小麦已被大量应用于小麦分子标记工作中。但是,目前还缺乏人工合成小麦的异源六倍化过程对微卫星影响的研究。本研究直接比较了四倍体小麦与节节麦远缘杂交并经染色体加倍获得人工合成小麦前后,位于普通小麦A/B染色体组不同染色体臂上的66个特异引物揭示的微卫星位点的保守性和可转移性。结果表明,除了一个引物在新合成小麦中扩增出供体亲本没有的新带,一个引物在节节麦扩增出的产物在新合成小麦中消失,其他的所有微卫星引物的扩增产物在小麦合成前后是保守的,没有变异发生。所有的引物能够在四倍体小麦中扩增出微卫星产物,四倍体小麦中的扩增产物也出现在新的人工合成小麦中;有70%的引物能够在节节麦扩增出产物,其中的绝大多数产物也出现在新的人工合成小麦中。因此,普通小麦A/B染色体组的这些微卫星引物除了在人工合成小麦的A/B染色体组中扩增出产物,还能在其D染色体组中扩增出产物,也就是说,这些引物对人工合成小麦而言,并非是A/B染色体组特异的。根据该研究结果,讨论了小麦微卫星的可转移性和特异性问题,重点讨论了在应用人工合成小麦构建的遗传群体进行微卫星分子标记中的应用价值及其应该注意的问题。     

人工合成小麦抗白粉病未知基因的SSR标记

YAV-2/TEZ//A.SQ(895)是硬粒小麦与粗山羊草杂交获得的抗白粉病人工合成小麦。本研究利用人工合成小麦YAV-2/TEZ//A.SQ(895)与感白粉病的普通小麦品系品资50098杂交和自交获得的F2代群体及F3家系,在温室条件下鉴定群体的白粉病抗性。遗传分析结果表明,该抗白粉病基因为显性单基因遗传。利用647对小麦SSR引物进行了白粉病抗性基因的分子标记分析,结果表明该白粉病抗性基因与2A染色体的6个SSR标记连锁,与标记Xcfa2086的遗传距离最近,为11.8cM。     

Genome-Wide Association Mapping for Resistance to Leaf and Stripe Rust in Winter-Habit Hexaploid Wheat Landraces

Leaf rust, caused by Puccinia triticina (Pt), and stripe rust, caused by P. striiformis f. sp. tritici (Pst), are destructive foliar diseases of wheat worldwide. Breeding for disease resistance is the preferred strategy of managing both diseases. The continued emergence of new races of Pt and Pst requires a constant search for new sources of resistance. Here we report a genome-wide association analysis of 567 winter wheat (Triticum aestivum) landrace accessions using the Infinium iSelect 9K wheat SNP array to identify loci associated with seedling resistance to five races of Pt (MDCL, MFPS, THBL, TDBG, and TBDJ) and one race of Pst (PSTv-37) frequently found in the Northern Great Plains of the United States. Mixed linear models identified 65 and eight significant markers associated with leaf rust and stripe rust, respectively. Further, we identified 31 and three QTL associated with resistance to Pt and Pst, respectively. Eleven QTL, identified on chromosomes 3A, 4A, 5A, and 6D, are previously unknown for leaf rust resistance in T. aestivum.     

Comparative High-Resolution Mapping of the Wax Inhibitors Iw1 and Iw2 in Hexaploid Wheat

The wax (glaucousness) on wheat leaves and stems is mainly controlled by two sets of genes: glaucousness loci (W1 and W2) and non-glaucousness loci (Iw1 and Iw2). The non-glaucousness (Iw) loci act as inhibitors of the glaucousness loci (W). High-resolution comparative genetic linkage maps of the wax inhibitors Iw1 originating from Triticum dicoccoides, and Iw2 from Aegilops tauschii were developed by comparative genomics analyses of Brachypodium, sorghum and rice genomic sequences corresponding to the syntenic regions of the Iw loci in wheat. Eleven Iw1 and eight Iw2 linked EST markers were developed and mapped to linkage maps on the distal regions of chromosomes 2BS and 2DS, respectively. The Iw1 locus mapped within a 0.96 cM interval flanked by the {"type":"entrez-nucleotide","attrs":{"text":"BE498358","term_id":"9696891","term_text":"BE498358"}}BE498358 and {"type":"entrez-nucleotide","attrs":{"text":"CA499581","term_id":"24990541","term_text":"CA499581"}}CA499581 EST markers that are collinear with 122 kb, 202 kb, and 466 kb genomic regions in the Brachypodium 5S chromosome, the sorghum 6S chromosome and the rice 4S chromosome, respectively. The Iw2 locus was located in a 4.1 to 5.4-cM interval in chromosome 2DS that is flanked by the {"type":"entrez-nucleotide","attrs":{"text":"CJ886319","term_id":"143584862","term_text":"CJ886319"}}CJ886319 and {"type":"entrez-nucleotide","attrs":{"text":"CJ519831","term_id":"93083902","term_text":"CJ519831"}}CJ519831 EST markers, and this region is collinear with a 2.3 cM region spanning the Iw1 locus on chromosome 2BS. Both Iw1 and Iw2 co-segregated with the {"type":"entrez-nucleotide","attrs":{"text":"BF474014","term_id":"11543196","term_text":"BF474014"}}BF474014 and {"type":"entrez-nucleotide","attrs":{"text":"CJ876545","term_id":"143556664","term_text":"CJ876545"}}CJ876545 EST markers, indicating they are most likely orthologs on 2BS and 2DS. These high-resolution maps can serve as a framework for chromosome landing, physical mapping and map-based cloning of the wax inhibitors in wheat.     

从CIMMYT引进的人工合成六倍体小麦D染色体组微卫星分子标记的遗传差异

采用微卫星（SSR）分子标记技术,选用23个D染色体组特异性引的对来自CIMMYT的26份人工合成六倍体小麦D染色体组的遗传多样性进行了分析。研究发现,26份材料在D染色体组上存在丰富的等位基因变异（92个）,平均每个基因座为4个。遗传距离计算结果也显示,26份材料D染色体组之间具有较大的遗传差异,平均遗传距离高达0．4955。因此,人工合成六倍体小麦D染色体组中存在丰富的遗传多样性,可以作为拓宽普通小麦遗传基础的新的遗传变异来源。研究还发现,由同一个粗山羊草基因型与不同硬粒小麦杂交合成的人工合成六倍体小麦（如合成种17和18）在所用检测的23个基因座中有3个存在差异,说明小麦在多倍化后,供体基因组在重复序列区域会发生遗传分化。     

小麦吸收土壤磷转运子在酵母突变体中的功能互补分析

以小麦磷转运子全长编码cDMA(TaPT2)为探针与小麦基因组DNA进行Southern杂交,结果表明,在小麦基因组中存在该基因的不同家族成员,另外,将TaPT2基因转入酵母突变体MB192中,以野生型菌株YPH084为对照,分别检测YTaPT2,YPH084和MB192酸性磷酸酶分泌情况,生长情况以及对培养基的磷吸收情况,得到结论：TaPT2的功能与酵母磷转运子编码基因PHO84相似,具有增强酵母吸收磷的作用。     

Genome-Wide Association Mapping for Seedling and Adult Plant Resistance to Stripe Rust in Synthetic Hexaploid Wheat

Use of genetic diversity from related wild and domesticated species has made a significant contribution to improving wheat productivity. Synthetic hexaploid wheats (SHWs) exhibit natural genetic variation for resistance and/or tolerance to biotic and abiotic stresses. Stripe rust caused by (Puccinia striiformis f. sp. tritici; Pst), is an important disease of wheat worldwide. To characterise loci conferring resistance to stripe rust in SHWs, we conducted a genome-wide association study (GWAS) with a panel of 181 SHWs using the wheat 9K SNP iSelect array. The SHWs were evaluated for their response to the prevailing races of Pst at the seedling and adult plant stages, the latter in replicated field trials at two sites in Ethiopia in 2011. About 28% of the SHWs exhibited immunity at the seedling stage while 56% and 83% were resistant to Pst at the adult plant stage at Meraro and Arsi Robe, respectively. A total of 27 SNPs in nine genomic regions (1BS, 2AS, 2BL, 3BL, 3DL, 5A, 5BL, 6DS and 7A) were linked with resistance to Pst at the seedling stage, while 38 SNPs on 18 genomic regions were associated with resistance at the adult plant stage. Six genomic regions were commonly detected at both locations using a mixed linear model corrected for population structure, kinship relatedness and adjusted for false discovery rate (FDR). The loci on chromosome regions 1AS, 3DL, 6DS and 7AL appeared to be novel QTL; our results confirm that resynthesized wheat involving its progenitor species is a rich source of new stripe (yellow) rust resistance that may be useful in choosing SHWs and incorporating diverse yellow rust (YR) resistance loci into locally adapted wheat cultivars.     

Cultivars of Hexaploid Wheat of Contrasting Stature and Chlorophyll Retention Differ in Cytokinin Content and Responsiveness

The work reported here compared cytokinin content and sensitivityin a selection of hexaploid wheat (Triticum aestivum L.) cultivarsusing the following measurements: leaf cytokinins at three timepoints during light-growth and at four 24 h intervals afterlight-grown plants were transferred to darkness; sensitivityof root growth to direct applications of isopentenyl adenosine([9R]iP); and, sensitivity of germination and subsequent rootand shoot growth to 18 h imbibition of seeds in benzyladenine(BA). Accumulation of zeatin riboside-type cultivars was greatestduring light-growth in Tibet Dwarf, a wheat with an extremedwarf phenotype, intermediate in Omar standard and dwarf cultivars,and lowest in the standard and dwarf versions of Itana. Cytokininlevels were otherwise not directly correlated to plant staturein these wheats. There were no cultivar-associated qualitativedifferences in the types of cytokinins detected in this study.During the 16 h light period, the content of zeatin riboside-typecytokinins increased up to tenfold and then declined to basallevels during dark growth. Chlorophyll retention during dark-growthwas correlated with leaf cytokinin content. Data collected ata restricted number of sampling points during dark-growth suggesteda cyclic accumulation of [9R]iP-type cytokinins and the apparentcycle in Tibet Dwarf was offset by 24 h. Tibet Dwarf showedthe greatest root growth inhibition after exposure of seedlingroots to [9R]iP or imbibition of seeds in BA. Neither of thesetreatments affected shoot growth in any of the cultivars. Wheat; Triticum aestivum ; cytokinin; zeatin riboside; benzyladenine; root inhibition     

Novel Genetic Variants of GA-Insensitive Rht-1 Genes in Hexaploid Wheat and Their Potential Agronomic Value

This study has found numerous novel genetic variants of GA-insensitive dwarfing genes with potential agricultural value for crop improvement. The cultivar, Spica is a tall genotype and possesses the wild-type genes of Rht-A1a, Rht-B1a and Rht-D1a. The cultivar Quarrion possesses a null mutant in the DELLA motif in each of the 3 genomes. This is a first report of a null mutant of Rht-A1. In addition, novel null mutants which differ from reported null alleles of Rht-B1b, Rht-B1e and Rht-D1b have been found in Quarrion, Carnamah and Whistler. The accession, Aus1408 has an allele of Rht-B1 with a mutation in the conserved ‘TVHYNP’ N-terminal signal binding domain with possible implications on its sensitivity to GA. Mutations in the conserved C-terminal GRAS domain of Rht-A1 alleles with possible effects on expression have been found in WW1842, Quarrion and Drysdale. Genetic variants with putative spliceosomal introns in the GRAS domain have been found in all accessions except Spica. Genome-specific cis-sequences about 124 bp upstream of the start codon of the Rht-1 gene have been identified for each of the three genomes.