A candidate recombination modifier gene for Zea mays L

Maize meiotic mutant desynaptic (dy) was tested as a candidate recombination modifier gene because its effect is manifested in prophase I. Recombination rates for desynaptic (dy) and its wild type were compared in two ways: (1) segregation analysis using six linked molecular markers on chromosome 1L and (2) cytogenetic analysis using fluorescence in situ hybridization (FISH)-aided meiotic configurations observed in metaphase I. Chromosome 1L map lengths among the six linked markers were 45-63 cM for five F2 dy/dy plants, significantly lower than the wild-type F2 map distance of 72 cM. Chromosomes 2 and 6 were marked with rDNA FISH probes, and their map lengths were estimated from FISH-adorned meiotic configurations using the expectation-maximization algorithm. Chiasma frequencies for dy/dy plants were significantly reduced for both arms of chromosome 2, for chromosome arm 6L, and for eight unidentified chromosomes. There was a notable exception for the nucleolus-organizing region-bearing arm chromosome arm 6S, where dy increased chiasma frequency. Maize meiotic mutant desynaptic is a recombination modifier gene based on cytogenetic and segregation analyses.     

Chromosome-specific molecular organization of maize (Zea mays L.) centromeric regions

A set of oat-maize chromosome addition lines with individual maize (Zea mays L.) chromosomes present in plants with a complete oat (Avena sativa L.) chromosome complement provides a unique opportunity to analyze the organization of centromeric regions of each maize chromosome. A DNA sequence, MCS1a, described previously as a maize centromere-associated sequence, was used as a probe to isolate cosmid clones from a genomic library made of DNA purified from a maize chromosome 9 addition line. Analysis of six cosmid clones containing centromeric DNA segments revealed a complex organization. The MCS1a sequence was found to comprise a portion of the long terminal repeats of a retrotransposon-like repeated element, termed CentA. Two of the six cosmid clones contained regions composed of a newly identified family of tandem repeats, termed CentC. Copies of CentA and tandem arrays of CentC are interspersed with other repetitive elements, including the previously identified maize retroelements Huck and Prem2. Fluorescence in situ hybridization revealed that CentC and CentA elements are limited to the centromeric region of each maize chromosome. The retroelements Huck and Prem2 are dispersed along all maize chromosomes, although Huck elements are present in an increased concentration around centromeric regions. Significant variation in the size of the blocks of CentC and in the copy number of CentA elements, as well as restriction fragment length variations were detected within the centromeric region of each maize chromosome studied. The different proportions and arrangements of these elements and likely others provide each centromeric region with a unique overall structure.     

Interchromosomal recombination in Zea mays

A new allele of the 27-kD zein locus in maize has been generated by interchromosomal recombination between chromosomes of two different inbred lines. A continuous patch of at least 11,817 bp of inbred W64A, containing the previously characterized Ra allele of the 27-kD zein gene, has been inserted into the genome of A188 by a single crossover. While both junction sequences are conserved, sequences of the two homologs between these junctions differ considerably. W64A contains the 7313-bp-long retrotransposon, Zeon-1. A188 contains a second copy of the 27-kD zein gene and a 2-kb repetitive element. Therefore, recombination results in a 7.3-kb insertion and a 14-kb deletion compared to the original S+A188 allele. If nonpairing sequences are looped out, 206 single base changes, frequently clustered, are present. The structure of this allele may explain how a recently discovered example of somatic recombination occurred in an A188/W64A hybrid. This would indicate that despite these sequence differences, pairing between these alleles could occur early during plant development. Therefore, such a somatically derived chimeric chromosome can also be heritable and give rise to new alleles.     

High efficiency transformation of maize (Zea mays L.) mediated by Agrobacterium tumefaciens

Transformants of maize inbred A188 were efficiently produced from immature embryos cocultivated with Agrobacterium tumefaciens that carried "super-binary" vectors. Frequencies of transformation (independent transgenic plants/embryos) were between 5% and 30%. Almost all transformants were normal in morphology, and more than 70% were fertile. Stable integration, expression, and inheritance of the transgenes were confirmed by molecular and genetic analysis. Between one and three copies of the transgenes were integrated with little rearrangement, and the boundaries of T-DNA were similar to those in transgenic dicotyledons and rice. F1 hybrids between A188 and five other inbreds were transformed at low frequencies.     

Cloning and biochemical characterization of an anionic peroxidase from Zea mays

We have isolated, cloned and characterized a cDNA from Zea mays L., denoted ZmAP1, coding for an anionic peroxidase. The open reading frame of ZmAP1 starting 72 residues from the 5' end of the cDNA predicts a 37,778 dalton protein of 356 amino acid residues. The protein has high similarity to other peroxidases and contains two peroxidase motifs that carry two highly conserved histidines in the active center. We expressed recombinant ZmAP1 protein in E. coli as a fusion with maltose-binding protein. The fusion protein was biochemically active after addition of hemin to the apoprotein. The maize peroxidase ZmAP1 has a pH optimum at pH 4.0 and a Km of 0.2 mM for the substrate 2,2'-azino-bis-(3-ethyl-benzothiazolin-6-sulfonic acid) at this pH. In maize seedlings the ZmAP1 gene is expressed predominantly in roots, the mesocotyl, the coleoptile and to a lower extent in the node, whereas no expression in the primary leaf was found. In situ hybridization shows that the expression of ZmAP1 in the young maize root is confined to the epidermis, hypodermis and the pericycle.     

芘胁迫对玉米幼苗生长及生理指标的影响

[目的]探讨芘胁迫对玉米幼苗生长及生理指标的影响.[方法]通过水培试验,研究了4个浓度(0、0.5、1.0和2.0 mg/L)的芘对2种玉米市祥1号(敏感品种)和广甜3号(耐性品种)幼苗生长及生理指标的影响.[结果]在试验浓度范围内,随芘浓度的升高,市祥1号叶绿素a和叶绿素b含量呈先升高后降低的趋势,相对生长量和绝对生长速率呈下降趋势,根系和叶片丙二醛(MDA)含量呈上升趋势;广甜3号相对生长量、绝对生长速率和叶绿素a含量呈先上升后下降趋势,叶绿素b和根系MDA含量呈上升趋势,叶片MDA含量呈下降趋势.与对照相比,市祥1号在0.5～1.0 mg/L芘处理对叶绿素a和叶绿素b含量显著增加,芘浓度≥2.0 mg/L时则显著降低,芘浓度≥1.0 mg/L时根系和叶片MDA含量显著增加,而相对生长量和绝对生长速率显著降低;广甜3号在芘浓度≥1.0 mg/L时叶绿素a和叶绿素b含量显著增加.芘浓度与敏感品种市祥1号的相对生长量和绝对生长速率呈显著负相关,与根系MDA含量呈显著正相关;芘浓度与耐性品种广甜3号叶绿素b含量和根系MDA含量呈显著正相关.[结论]为玉米苗期的管理措施提供了依据.     

Genetic mechanisms underlying apimaysin and maysin synthesis and corn earworm antibiosis in maize (Zea mays L.)

C-glycosyl flavones in maize silks confer resistance (i.e., antibiosis) to corn earworm (Helicoverpa zea [Boddie]) larvae and are distinguished by their B-ring substitutions, with maysin and apimaysin being the di- and monohydroxy B-ring forms, respectively. Herein, we examine the genetic mechanisms underlying the synthesis of maysin and apimaysin and the corresponding effects on corn earworm larval growth. Using an F2 population, we found a quantitative trait locus (QTL), rem1, which accounted for 55.3% of the phenotypic variance for maysin, and a QTL, pr1, which explained 64.7% of the phenotypic variance for apimaysin. The maysin QTL did not affect apimaysin synthesis, and the apimaysin QTL did not affect maysin synthesis, suggesting that the synthesis of these closely related compounds occurs independently. The two QTLs, rem1 and pr1, were involved in a significant epistatic interaction for total flavones, suggesting that a ceiling exists governing the total possible amount of C-glycosyl flavone. The maysin and apimaysin QTLs were significant QTLs for corn earworm antibiosis, accounting for 14. 1% (rem1) and 14.7% (pr1) of the phenotypic variation. An additional QTL, represented by umc85 on the short arm of chromosome 6, affected antibiosis (R2 = 15.2%), but did not affect the synthesis of the C-glycosyl flavones.     

A modified autonomous En transposon in maize (Zea mays L.) elicits a differential response of reporter alleles

Transposable elements in maize are composed of a defined molecular structure that includes coding sequences, determiners of functionality and ordered terminal motifs that provide binding sites for transposase proteins. Alterations in these components change the phenotypic expression of unstable genes with transposon inserts. The molecular basis for the altered timing and frequency of transposition as determined by the size and number of spots on kernels or stripes on leaves has generally been described for defective inserts in genes. Most differential patterns can be ascribed to alterations in the terminal motifs of the reporter allele structure that supplies a substrate (terminal inverted repeat motifs) for transposase activity. For autonomously functioning alleles, the explanations for changes in phenotype are not so clear. In this report, an En-related element identified as F-En is described that shares with En the recognition of a specific defective element cl(mr)888104 but differs from En in that this F-En element does not recognize the canonical cl(mr) elements that are recognized by En. Evidence is provided suggesting that F-En does not recognize other En/Spm-related defective elements, some of whose sequences are known. This modified En arose from a cl-m autonomously mutating En allele.     

微针注射甜玉米胚芽gus基因转化瞬时表达研究

[目的]研究微针注射甜玉米(Zea mays L.)胚芽gus基因转化瞬时表达,为玉米遗传转化的进一步研究奠定基础.[方法]以农杆菌为介导,采用gus基因瞬时表达技术,建立了胚芽注射开放性基因转化体系.[结果]3 mm长的胚芽为最适转化受体;农杆菌EHA105菌液浓度最适OD值为0.558～0.630;gus基因转化最适共培养天数为3 d.在以上最适转化条件下,微针注射胚芽生长点基因转化率为1%.[结论]该研究可为改良甜玉米遗传性状,培育基因工程新品种提供参考.     

Localization of photosynthetic electron transport components in mesophyll and bundle sheath chloroplasts of Zea mays

The organization of the electron transport components in mesophyll and bundle sheath chloroplasts of Zea mays was investigated. Grana-containing mesophyll chloroplasts (chlorophyll a to chlorophyll b ratio of about 3.0) possessed the full complement of the various electron transport components, comparable to chloroplasts from C3 plants. Agranal bundle sheath chloroplasts (Chl a/Chl b greater than 5.0) contained the full complement of photosystem (PS) I and of cytochrome (cyt) f but lacked a major portion of PS II and its associated Chl a/b light-harvesting complex (LHC), and most of the cyt b559. The kinetic analysis of system I photoactivity revealed that the functional photosynthetic unit size of PS I was unchanged and identical in mesophyll and bundle sheath chloroplasts. The results suggest that PS I is contained in stroma-exposed thylakoids and that it does not receive excitation energy from the Chl a/b LHC present in the grana. A stoichiometric parity between PS I and cyt f in mesophyll and bundle sheath chloroplasts indicates that biosynthetic and functional properties of cyt f and P700 are closely coordinated. Thus, it is likely that both cyt f and P700 are located in the membrane of the intergrana thylakoids only. The kinetic analysis of PS II photoactivity revealed the absence of PS II alpha from the bundle sheath chloroplasts and helped identify the small complement of system II in bundle sheath chloroplasts as PS II beta. The distribution of the main electron transport components in grana and stroma thylakoids is presented in a model of the higher plant chloroplast membrane system.     

Relationship of senescence of corn (Zea mays L.) stalk pith, cob parenchyma and first developed leaf tissues to RNA content and synthesis

In stalk parenchyma tissue of WF9 X 38-11 single corn (Zea mays L.), there was a per cell increase in RNA synthesis and a slight increase in total RNA as cells became older. In cob parenchyma tissue of WF9 X 38-11 single cross corn, both total RNA content and RNA synthesis per cell decreased with the age of cells. In first developed leaf tissue of FR43 X FR14A single cross corn, RNA synthesis increased steadily but only slightly over its life span. The pattern of RNA synthesis and destruction in senescing leaf tissue of seedlings and two sources of parenchyma tissue of maturing plants appeared to differ.     

Cloning of two cDNAs encoding calnexin-like and calreticulin-like proteins from maize (Zea mays) leaves: identification of potential calcium-binding domains

The U-turn (uridine turn) is an RNA structural motif that contains a change in backbone direction stabilized by specific interactions across the bend. It was first identified in the anticodon loop and the T-loop of yeast tRNA(Phe) (Quigley & Rich, 1976, Science 194:796-806) and has recently also been found in the crystal structure of the hammerhead ribozyme (Pley HW, Flaherty KM, McKay DB, 1994a, Nature 372:68-74). These U-turn motifs follow a UNR consensus sequence (where N is any nucleotide and R is G or A). Here we report that the frequently occurring GNRA tetraloops also contain a U-turn motif, and we discuss the role of U-turns as abundant tertiary structural motifs in RNA.     

紫茎泽兰不同组织水提取液对水稻·玉米化感作用机制

[目的]研究紫茎泽兰不同组织根、茎、叶水浸提取液对2种农作物玉米、水稻化感作用及对受试农作物生理指标的影响,以探讨化感物质作用机制.[方法]采用培养皿滤纸法测定不同浓度根、茎、叶水浸提液对受试种子萌发率和幼苗生长影响,并检测受试农作物生理指标丙二醛(MDA)含量、过氧化物酶(POD)活力的变化.[结果]紫茎泽兰根、茎、叶水浸提液对受试玉米、水稻的发芽率和幼苗根长、株高均有不同程度的影响,使受试农作物幼苗体内丙二醛(MDA)含量增加、过氧化物酶(POD)活性呈现先增加后降低趋势.[结论]紫茎泽兰不同组织水提液对受体植物的种子萌发、幼苗生长过程均具有明显的抑制作用,其茎的化感敏感效应大于根、叶,水提液抑制受试农作物光合效率,提高了幼苗体内的MDA含量,并降低了POD酶活性.