Identification of the genomic locations of duplicate nucleotide sequences in maize by analysis of restriction fragment length polymorphisms

While preparing a linkage map for maize based upon loci detected through the use of restriction fragment length polymorphisms (RFLPs), it was found that 62 of the 217 cloned maize sequences tested (29%) detected more than one fragment on genomic Southern blots. Thus, more than one nucleotide sequence is present within the maize genome which is in part homologous to each of these cloned sequences. The genomic locations of these ``duplicate' sequences were determined and it was found that they usually originated from different chromosomes. The process which produced them did not operate randomly as some pairs of chromosomes share many duplicate sequences while many other pairs share none. Furthermore, these shared duplicate sequences are generally arrayed in an ordered arrangement along these chromosomes. It is believed that chromosomal segments which contain several duplicate loci in a generally ordered arrangement must have had a common origin. The presence of these duplicated segments supports the idea that allopolyploidy may have been involved in the evolution of maize. Nevertheless, the duplicate loci do not primarily involve five pairs of chromosomes and thus, five pairs of homeologous chromosomes are not currently present within the maize genome. The data clearly indicate that maize is not a recent allotetraploid produced by hybridization between two individuals with similar genomic structures; however, the data are also consistent with the possibility of these shared duplicate chromosomal segments having been generated through internal duplication.     

玉米雄性不育资源的发掘与利用

植物雄性不育是指植物雄性生殖器官不能产生正常有功能花粉的现象.玉米(Zea mays L.)是重要的粮食作物之一,也是较早利用杂种优势的作物之一.当前,生产上广泛种植的玉米品种类型主要是单交种.我国玉米杂交种的播种面积常年稳定在6.2亿亩左右,年用种量10亿公斤以上,常年制种面积高达250多万亩.利用传统的人工去雄或机...     

Hybridization of maize and teosinte,in mexico and guatemala and the improvement of maize

The recognition and subsequent detection of the importance of teosinte introgression in the racial diversity and heterotic gene architecture of maize has been one of the outstanding achievements of Paul C. Mangelsdorf’s investigations into the origin of maize. This paper documents three areas in Mexico and Guatemala where maize and teosinte hybridize and where there is a system by which native cultivators exploit the heterotic nature of maize to increase their harvest. There is little reason to doubt that the hybridization and subsequent introgression of teosinte genes into maize observed at these sites is changed from that which has occurred over the past three thousand years resulting in the tremendous diversity and pronounced hybrid vigor in maize.     

The Rh phenotype r'r' in Polynesians

Since 1970, 19 persons with the r'r' phenotype have been detected at the Auckland Blood Transfusion Centre. All have been Maoris or Pacific Islanders. This study reports the detailed serological investigations on 11 r'r' samples. No evidence was found for the presence of the Du antigen, and there was no evidence that the Polynesian r' was related to the Negroid r'. It is postulated that the r' (Cde) of Polynesians arose from a mutation of CDe. No morphological abnormalities of r'r' red cells were found. Blood samples were also tested for various high- and low-frequency antigens, and one specimen was found to be r'r' Jk(a-b-).     

Following Tetraploidy in Maize,a Short Deletion Mechanism Removed Genes Preferentially from One of the Two Homeologs

Previous work in Arabidopsis showed that after an ancient tetraploidy event, genes were preferentially removed from one of the two homeologs, a process known as fractionation. The mechanism of fractionation is unknown. We sought to determine whether such preferential, or biased, fractionation exists in maize and, if so, whether a specific mechanism could be implicated in this process. We studied the process of fractionation using two recently sequenced grass species: sorghum and maize. The maize lineage has experienced a tetraploidy since its divergence from sorghum approximately 12 million years ago, and fragments of many knocked-out genes retain enough sequence similarity to be easily identifiable. Using sorghum exons as the query sequence, we studied the fate of both orthologous genes in maize following the maize tetraploidy. We show that genes are predominantly lost, not relocated, and that single-gene loss by deletion is the rule. Based on comparisons with orthologous sorghum and rice genes, we also infer that the sequences present before the deletion events were flanked by short direct repeats, a signature of intra-chromosomal recombination. Evidence of this deletion mechanism is found 2.3 times more frequently on one of the maize homeologs, consistent with earlier observations of biased fractionation. The over-fractionated homeolog is also a greater than 3-fold better target for transposon removal, but does not have an observably higher synonymous base substitution rate, nor could we find differentially placed methylation domains. We conclude that fractionation is indeed biased in maize and that intra-chromosomal or possibly a similar illegitimate recombination is the primary mechanism by which fractionation occurs. The mechanism of intra-chromosomal recombination explains the observed bias in both gene and transposon loss in the maize lineage. The existence of fractionation bias demonstrates that the frequency of deletion is modulated. Among the evolutionary benefits of this deletion/fractionation mechanism is bulk DNA removal and the generation of novel combinations of regulatory sequences and coding regions.     

玉米花药培养和单倍体育种的研究新进展

利用花药培养获得单倍体,从而加速育种进程,是一顶新兴的生物技术,目前在玉米育种中广泛应用,本文综合近几年来国内外玉米的花药培养、单位体育种以及基因工程等方面的研究进展,重点对影响玉米花药培养效率的诸多因素进行了详细论述,并讨论了利用单7保体植株进行基因转导的潜力。     

Proteome analysis of maize roots reveals that oxidative stress is a main contributing factor to plant arsenic toxicity

To gain insight into plant responses to arsenic, the effect of arsenic exposure on maize (Zea mays L.) root proteome has been examined. Maize seedlings were fed hydroponically with 300 microM sodium arsenate or 250 microM sodium arsenite for 24 h, and changes in differentially displayed proteins were studied by two-dimensional electrophoresis and digital image analysis. About 10% of total detected maize root proteins (67 out of 700) were up- or down-regulated by arsenic, among which 20 were selected as being quite reproducibly affected by the metalloid. These were analyzed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and 11 of them could be identified by comparing their peptide mass fingerprints against protein- and expressed sequence tag-databases. The set of identified maize root proteins highly responsive to arsenic exposure included a major and functionally homogeneous group of seven enzymes involved in cellular homeostasis for redox perturbation (e.g., three superoxide dismutases, two glutathione peroxidases, one peroxiredoxin, and one p-benzoquinone reductase) besides four additional, functionally heterogeneous, proteins (e.g., ATP synthase, succinyl-CoA synthetase, cytochrome P450 and guanine nucleotide-binding protein beta subunit). These findings strongly suggest that the induction of oxidative stress is a main process underlying arsenic toxicity in plants.     

玉米花药培养和单倍体育种的研究新进展

利用花药培养获得单倍体,从而加速育种进程,是一项新兴的生物技术,目前在玉米育种中广泛应用。本文综合近几年来国内外玉米的花药培养、单倍体育种以及基因工程等方面的研究进展,重点对影响玉米花药培养效率的诸多因素进行了详细论述,并讨论了利用单倍体植株进行基因转导的潜力。     

Purification and partial characterization of an acidic ribosomal protein kinase from maize

Phosphorylation and dephosphorylation of ribosomal proteins have been suggested to participate in the regulation of protein synthesis in eukaryotic organisms. The present research focuses on the purification and partial characterization of a protein kinase from maize ribosomes that specifically phosphorylates acidic ribosomal proteins. Ribosomes purified from maize axes were used as the enzyme source. Purification of ribosomes was performed by centrifugation through a 0.5 M sucrose, 0.8 M KCl cushion. A protein kinase activity present in this fraction was released by extraction with 1.5 M KCl and further purified by diethylaminoethyl cellulose column chromatography. A peak containing protein kinase activity was eluted around 400 m M KCl. Analysis of this fraction by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed one band of 38 kDa molecular mass, which cross-reacted in a western blot with antibodies raised against proteins from the large ribosomal subunit. This enzyme specifically phosphorylates one of the acidic ribosomal proteins (P₂). Its activity is inhibited by Ca²⁺ and Zn²⁺ and is activated by Mg²⁺, polylysine and spermine. The relevance of this protein kinase in reinitiating the protein synthesis process during germination is discussed.     

转人工miRNA抗玉米粗缩病毒载体玉米的培育及其抗病能力

玉米是重要的粮食作物,水稻黑条矮缩病毒(RBSDV)是玉米粗缩病的病原,由其引起的玉米粗缩病给玉米生产造成重大损失。利用人工mi RNA构建抗病毒植物的技术已经在多种植物中被证明有效,但是在玉米中的尝试未见报道。实验根据玉米zea-mi R159a的前体序列和RBSDV基因组中编码功能蛋白的基因和基因沉默抑制子的序列信息设计引物,构建了用于沉默RBSDV编码基因和基因沉默抑制子的ami RNA(Artificial mi RNA)基因。构建p CAMBIA3301-121-ami RNA植物表达载体,利用农杆菌介导法转化玉米自交系综31(Z31)。对转基因玉米进行分子检测,选择mi RNA表达量高的纯合体株系进行自然发病实验,按0-4的分级标准调查玉米粗缩病的严重度。结果表明,转抗粗缩病毒人工mi RNA载体玉米纯合体株系的抗病表现好于野生型玉米,其中针对基因组6的S6-mi R159转基因玉米抗病情况较好。研究表明利用人工mi RNA技术构建抗病毒病玉米新品种是可行的。     

Utility of different gene enrichment approaches toward identifying and sequencing the maize gene space

Maize (Zea mays) possesses a large, highly repetitive genome, and subsequently a number of reduced-representation sequencing approaches have been used to try and enrich for gene space while eluding difficulties associated with repetitive DNA. This article documents the ability of publicly available maize expressed sequence tag and Genome Survey Sequences (GSSs; many of which were isolated through the use of reduced representation techniques) to recognize and provide coverage of 78 maize full-length cDNAs (FLCs). All 78 FLCs in the dataset were identified by at least three GSSs, indicating that the majority of maize genes have been identified by at least one currently available GSS. Both methyl-filtration and high-Cot enrichment methods provided a 7- to 8-fold increase in gene discovery rates as compared to random sequencing. The available maize GSSs aligned to 75% of the FLC nucleotides used to perform searches, while the expressed sequence tag sequences aligned to 73% of the nucleotides. Our data suggest that at least approximately 95% of maize genes have been tagged by at least one GSS. While the GSSs are very effective for gene identification, relatively few (18%) of the FLCs are completely represented by GSSs. Analysis of the overlap of coverage and bias due to position within a gene suggest that RescueMu, methyl-filtration, and high-Cot methods are at least partially nonredundant.     

An Asian-specific 9-bp deletion of mitochondrial DNA is frequently found in Polynesians.

One hundred fifty Polynesians from five different island groups (Samoans, Maoris, Niueans, Cook Islanders, and Tongans) were surveyed for the presence of an Asian-specific length mutation of mitochondrial (mt) DNA by using enzymatic amplification with thermostable Taq DNA polymerase. Ninety-three percent of Polynesians exhibited this 9-bp deletion, including 100% of Samoans, Maoris, and Niueans. The same deletion was also found in 8% of Tolais from New Britain and in 14% of coastal New Guineans. A deletion frequency of 82% in Fijians confirmed their ethnic affinity to Polynesians. In contrast, the deletion was absent in 30 New Guinea highlanders and 31 Australian aborigines, the only exception being an aborigine who also had the Southeast Asian triplicated zeta-globin gene rearrangement in his nuclear DNA. These data support the theories claiming that an independent group of pre-Polynesian ancestors who colonized into the Pacific were ultimately derived from east Asia.     

Molecular evidence for phytosiderophore‐induced improvement of iron nutrition of peanut intercropped with maize in calcareous soil

Peanut/maize intercropping is a sustainable and effective agroecosystem that evidently enhances the Fe nutrition of peanuts in calcareous soils. So far, the mechanism involved in this process has not been elucidated. In this study, we unravel the effects of phytosiderophores in improving Fe nutrition of intercropped peanuts in peanut/maize intercropping. The maize ys3 mutant, which cannot release phytosiderophores, did not improve Fe nutrition of peanut, whereas the maize ys1 mutant, which can release phytosiderophores, prevented Fe deficiency, indicating an important role of phytosiderophores in improving the Fe nutrition of intercropped peanut. Hydroponic experiments were performed to simplify the intercropping system, which revealed that phytosiderophores released by Fe‐deficient wheat promoted Fe acquisition in nearby peanuts and thus improved their Fe nutrition. Moreover, the phytosiderophore deoxymugineic acid (DMA) was detected in the roots of intercropped peanuts. The yellow stripe1‐like (YSL) family of genes, which are homologous to maize yellow stripe 1 (ZmYS1), were identified in peanut roots. Further characterization indicated that among five AhYSL genes, AhYSL1, which was localized in the epidermis of peanut roots, transported Fe(III)–DMA. These results imply that in alkaline soil, Fe(III)–DMA dissolved by maize might be absorbed directly by neighbouring peanuts in the peanut/maize intercropping system.     

The allotetraploidization of maize

Summary Allotetraploidization is the creation of artifical allotetraploids from a normally diploid species. The possible value of allotetraploid maize has been discussed in Section I of this series. Allotetraploidization of maize can be achieved by restructuring a maize genome so that its chromosomes will not pair with those of the standard maize genome. This restructuring can be done by concentrating differential pairing affinity (DPA) factors into a single line by a recurrent selection type of breeding program. Because the divergence of the maize genome is a gradual process, it is necessary to devise a model for chromosome pairing and gene segregation in segmental allotetraploids. This has been done by considering pairing in each arm separately and then combining paired arms to form pairing configurations for whole chromosomes. The chromosome disjunction patterns are hypothesized and genetic ratios in relation to different levels of DPA are suggested.Contribution from the Science and Education Administration, U.S. Department of Agriculture, and the Agronomy Department, University of Missouri, Columbia, Missouri, Agricultural Experiment Station Journal Series No. 8090     

Gamete formation via meiotic nuclear restitution generates fertile amphiploid F1 (oat?×?maize) plants

Hybrid (oat×maize) zygotes developed into euhaploid plants with complete oat chromosome complements without maize chromosomes and into aneuhaploid plants with complete oat chromosome complements and different numbers of retained individual maize chromosomes. The elimination of maize chromosomes in the hybrid embryo is caused by uniparental genome loss during early steps of embryogenesis. Some of these haploid plants set seed in up to 50% of their self-pollinated spikelets. The high fertility was found to be mainly caused by formation of numerically unreduced female and male gametes (nunreduced=3x+0…3=21…24 chromosomes). Gamete formation involves meiotic nuclear restitution. The restitution process is caused by an alternative type of meiosis. It follows the model of levigatum-type semi-heterotypic divisions, but with a formation of the nuclear membrane at the transition from telophase I to interkinesis, which resembles the model of pygaera-type pseudo-homotypic divisions. We propose the name haploid meiotic restitution for this particular process combination. We discuss the use and implications of the specific process of gamete formation in F1 (oat×maize) plants.     

The maize b1 paramutation control region causes epigenetic silencing in Drosophila melanogaster

Paramutation is an epigenetic process in which a combination of alleles in a heterozygous organism results in a meiotically stable change in expression of one of the alleles. The mechanisms underlying paramutation are being actively investigated, and examples have been described in both plants and mammals, suggesting that it may utilize epigenetic mechanisms that are widespread and evolutionarily conserved. Paramutation at the well-studied maize b1 locus requires a control region consisting of seven 853 bp tandem repeats. To study the conservation of the epigenetic mechanisms underlying seemingly unique epigenetic processes such as paramutation, we created transgenic Drosophila melanogaster carrying the maize b1 control region adjacent to the Drosophila white reporter gene. We show that the b1 tandem repeats cause silencing of the white reporter in Drosophila. A single copy of the tandem repeat sequence is sufficient to cause silencing, and silencing strength increases as the number of tandem repeats increases. Additionally, transgenic lines with the full seven tandem repeats demonstrate evidence of either pairing-sensitive silencing and silencing in trans, or epigenetic activation in trans. These trans-interactions are dependent on repeat number, similar to maize b1 paramutation. Also, as in maize, the tandem repeats are bidirectionally transcribed in Drosophila. These results indicate that the maize b1 tandem repeats function as an epigenetic silencer and mediate trans-interactions in Drosophila, and support the hypothesis that the mechanisms underlying such epigenetic processes are conserved.     

The sweet potato—Its origin and primitive storage practices

In pre-Columbian times sweet potatoes provided food for the widely separated Mayans of Central America, the Incas of Peru and the Maoris of New Zealand, while two other species of the genus were occasionally used by Indians of western North America.     

玉米穗腐病抗性鉴定、遗传分析与分子机制*

玉米是我国主要农作物之一,其产量占全国谷物总产量三分之一左右,在国民生活中发挥了重要作用。玉米生长过程中受多种病害侵扰,其中穗腐病是一种由真菌导致的常见病害,目前已鉴定出40余种可诱发玉米穗腐病的病原菌。穗腐病不仅可造成玉米产量损失也导致籽粒品质严重下降,病原微生物所产生次生毒素更是危及人畜安全。目前穗腐病防治以化学方法为主,但是因此所造成的种植成本增加和环境污染问题日益突出,选育抗性品种成为防控玉米穗腐病最经济和安全有效的方法。玉米穗腐病抗性属于典型数量性状,国内外已有多个研究组通过建立玉米穗腐病抗性研究体系,开展玉米群体大规模穗腐病抗性鉴定工作,并筛选出一批抗病材料,这为玉米穗腐病抗性遗传改良奠定了材料基础。利用数量性状位点(quantitative trait locus, QTL)定位和全基因组关联分析(genome-wide association study, GWAS)等方法在玉米 1~10号染色体均检测到显著相关位点。尽管如此,玉米穗腐病抗性研究成果应用于生产的实例较少,生产上仍然缺乏综合性状优良且高抗穗腐病的玉米品种。这一方面是由于玉米穗腐病抗性遗传机制十分复杂、抗性基因克隆工作进展缓慢;另一方面为缺乏对玉米穗腐病抗性遗传研究进展进行系统总结,未有效开发分子标记用于分子育种所致。通过综述玉米穗腐病抗性遗传研究进展,汇总已定位QTL位点和显著关联SNP,构建一致性图谱和鉴定出定位热点区间,并进一步对比分析定位区间候选基因特征和转录组、代谢组研究进展,对促进玉米穗腐病抗性机制研究和玉米抗性育种均具有重要意义。