小麦Kr基因在小麦与玉米或鸭茅状摩擦禾杂交中的失活

用３７个小麦（Ｔｒｉｔｉｃｕｍａｅｓｔｉｖｕｍ）品种（系）为母本,分别与黑麦（Ｓｅｃａｌｅｃｅｒｅａｌｅ）、球茎大麦（Ｈｏｒｄｅｕｍｂｕｌｂｏｓｕｍ）、玉米（Ｚｅａｍａｙｓ）和鸭茅状摩擦禾（Ｔｒｉｐｓａｃｕｍｄａｃｔｙｌｏｉｄｅｓ）杂交,比较其亲和性,小麦和玉米或鸭茅状摩擦禾杂交比小麦与黑麦或球茎大麦杂交的亲和性显著提高。携带着显性Ｋｒ１和Ｋｒ２基因的小麦品种Ｈｏｐｅ与黑麦杂交,不能形成胚,而与玉米及鸭茅状摩擦禾杂交时,成胚率分别达１６．００％和３２．５０％。表明控制小麦与黑麦及球茎大麦杂交亲和性的Ｋｒ基因系统在小麦与玉米及小麦与鸭茅状摩擦禾属间杂交中失活。讨论了还存在有其它控制小麦属间杂交亲和性的遗传调控系统的可能性。     

Emb15 encodes a plastid ribosomal assembly factor essential for embryogenesis in maize

Ribosome assembly factors guide the complex process by which ribosomal proteins and the ribosomal RNAs form a functional ribosome. However, the assembly of plant plastid ribosomes is poorly understood. In the present study, we discovered a maize (Zea mays) plastid ribosome assembly factor based on our characterization of the embryo defective 15 (emb15) mutant. Loss of function of Emb15 retards embryo development at an early stage, but does not substantially affect the endosperm, and causes an albino phenotype in other genetic backgrounds. EMB15 localizes to plastids and possesses a ribosome maturation factor M (RimM) domain in the N-terminus and a predicted UDP-GlcNAc pyrophosphorylase domain in the C-terminus. The EMB15 RimM domain originated in bacteria and the UDP-GlcNAc pyrophosphorylase domain originated in fungi; these two domains came together in the ancestor of land plants during evolution. The N-terminus of EMB15 complemented the growth defect of an Escherichia coli strain with a RimM deletion and rescued the albino phenotype of emb15 homozygous mutants. The RimM domain mediates the interaction between EMB15 and the plastid ribosomal protein PRPS19. Plastid 16S rRNA maturation is also significantly impaired in emb15. These observations suggest that EMB15 functions in maize seed development as a plastid ribosome assembly factor, and the C-terminal domain is not important under normal conditions.     

Differentiation of restriction sites in ribosomal DNA in the genusApodemus

Southern blot analysis of ribosomal DNA (rDNA) from seven species ofApodemus was carried out in order to examine the genetic relationships between the species. Analysis of heterogeneity in rDNA spacers inA. sylvaticus, A. flavicollis, A. semotus, A. agrarius, A. argenteus, A. speciosus, andA. peninsulae, using 13 different restriction enzymes and cloned mouse rDNA probes, revealed that the families of rDNA in these species can be characterized by restriction maps which show the major constituents of rDNA repeating units (repetypes). Based on differences in the arrangement of restriction sites, sequence divergence among the different major repetypes was estimated. Among the seven species ofApodemus examined, the major repetypes ofA. flavicollis andA. sylvaticus were the most closely related, having only 1.0% sequence divergence. These repetypes and those of the remaining five species differ substantially from one another, with 4.3–8.5% divergence.This study was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science and Culture, Japan.     

A Cre::FLP fusion protein recombines FRT or loxP sites in transgenic maize plants

SUMMARY: The coding sequences of Cre (site-specific recombinase from bacteriophage P1) and FLP (yeast 2-microm plasmid site-specific recombinase) were fused in frame to produce a novel, dual-function, site-specific recombinase gene. Transgenic maize plants containing the Cre::FLP fusion expression vector were crossed to transgenic plants containing either the loxP or FRT excision substrate. Complete and precise excisions of chromosomal fragments flanked by the respective target sites were observed in the F1 and F2 progeny plants. The episomal DNA recombination products were frequently lost. Non-recombined FRT substrates found in the F1 plants were recovered in the F2 generation after the Cre::FLP gene segregated out. They produced the recombination products in the F3 generation when crossed back to the FLP-expressing plants. These observations may indicate that the efficiency of site-specific recombination is affected by the plant developmental stage, with site-specific recombination being more prevalent in developing embryos. The Cre::FLP fusion protein was also tested for excisions catalysed by Cre. Excisions were identified in the F1 plants and verified in the F2 plants by polymerase chain reaction and Southern blotting. Both components of the fusion protein (FLP and Cre) were functional and acted with similar efficiency. The crossing strategy proved to be suitable for the genetic engineering of maize using the FLP or Cre site-specific recombination system.     

Polysome formation and incorporation of new ribosomes into polysomes during germination of the embryonic axis of maize

Isolated axes of Zea mays L. cvs CiV₂ and CUZCO were imbibed for different periods of time, and free polysomes were extracted and analysed by centrifugation in a sucrose gradient. The amount of rRNA per axis was determined at different moments of germination. Polysome reassembly was practically completed by 8 h and 54% of the preformed ribosomes were found in the polysome fraction. An increase in the proportion of large polysomes was also observed during this period of germination. During the following period, the polysome content and the distribution of the various classes of polysomes remained unchanged.
The time of appearance of newly synthesized ribosomes into the polysomes was investigated using axes germinated in the presence of [³H]-uridine. Centrifugal analysis of EDTA-dissociated polysomes and gel electrophoretic analysis of polysomal RNA showed that new ribosomes appeared into polysomes a few hours after completion of the initial polysome assembly. When released into the cytoplasm, the new ribosomes were preferentially incorporated into polysomes rather than stored as free ribosomes.     

DNA endoreduplication in maize endosperm cells: the effect of exposure to short-term high temperature

DNA endoreduplication in Zea mays L. (cv. A619 × W64A) endosperm peaks between 16 and 18 d after pollination (DAP). The physiological function of DNA endoreduplication is not known but it is believed to be important in maize kernel development. In the present study, we investigated how 2, 4 or 6 d of high temperature (35 °C) affected DNA endoreduplication and maize kernel development in comparison with control kernels grown at 25 °C. Data were collected on fresh weight (FW), nuclei number, mitotic index, and DNA endoreduplication. Maize endosperm FW and nuclei number were reduced by exposure to 4 or 6 d of high temperature. At 18 DAP, the 2 d high temperature treatment (HTT) caused a reduction in FW and nuclei number, but had no effect on DNA endoreduplication and average DNA content per endosperm. However, when the exposure to high temperature was increased to 4 or 6 d, FW, nuclei number and the magnitude of DNA endoreduplication were progressively reduced, and the peak mitotic index was delayed compared with the control endosperm. At 18 DAP, the 4 d treatment showed 54·7% of the cells were 3 or 6 C, whereas only 41·2% were 12 C or higher. Six days of high temperature also resulted in a reduction in endosperm FW, nuclei number and a delay in the peak of mitotic index. DNA endoreduplication occurred in the kernels exposed to this treatment, although the magnitude was severely reduced compared with the control kernels. Nuclear DNA content was highly correlated (r = 0·93) with kernel FW, suggesting an important role of DNA endoreduplication in determining endosperm FW. The data suggest that high temperature during endosperm cell division exerted negative effects on DNA endoreduplication by dramatically reducing the nuclei number, leaving fewer nuclei available for DNA endoreduplication. However, the data also suggest that prolonged exposure to high temperature restricts entry of mitotic cells into the endoreduplication phase of the cell cycle.     

Homology between the ribosomal DNA of Escherichia coli and mitochondrial DNA preparations of maize is principally to sequences other than mitochondrial rRNA genes

E. coli ribosomal DNA has been used to probe maize mitochondrial DNA. It hybridizes primarily with chloroplast ribosomal DNA sequences and with fungal and bacterial sequences which may contaminate the mtDNA preparations. It also hybridizes to the chloroplast 16S ribosomal RNA gene sequence present in the mitochondrial genome (1) as well as to the mitochondrial 18S ribosomal RNA gene sequence. Weak sequence homology was detected between E. coli rDNA and the mitochondrial 26S ribosomal RNA gene.     

Differentiation of the endemic New Zealand greenheaded and brownheaded leafroller moths by restriction fragment length variation in the ribosomal gene complex

This study identified markers in the ribosomal RNA gene complex (rDNA) and evaluated their utility in the differentiation of four endemic New Zealand species of leafroller moth, Planotortrix excessana (Walker), P. octo (Dugdale), Ctenopseustis herana (Felder and Rogenhofer) and C. obliquana (Walker). Pooled genomic DNA from laboratory-reared specimens of C. herana and C. obliquana could be distinguished with a single restriction enzyme (e.g. ClaI), while P. excessana and P. octo could be satisfactorily distinguished using double digestions with either ClaI/PvuII or ClaI/BglII. It is concluded mat these markers may be used to identify moths from wild populations.     

Genome size and numerical polymorphism for the B chromosome in races of maize (Zea mays ssp. mays, Poaceae)

Twenty-one native populations (1120 individuals) of maize from Northern Argentina were studied. These populations, which belong to 13 native races, were cultivated at different altitudes (80-3620 m). Nineteen of the populations analyzed showed B chromosome (Bs) numerical polymorphism. The frequency of individuals with Bs varied from 0 to 94%. The number of Bs per plant varied from 0 to 8 Bs, with the predominant doses being 0, 1, 2, and 3. Those populations with varying number of Bs showed a positive and statistically significant correlation of mean number of Bs with altitude. The DNA content, in plants without Bs (A-DNA)(2n = 20), of 17 populations of the 21 studied was determined. A 36% variation (5.0-6.8 pg) in A-DNA content was found. A significant negative correlation between A-DNA content and altitude of cultivation and between A-DNA content and mean number of Bs was found. This indicates that there is a close interrelationship between the DNA content of A chromosomes and doses of Bs. These results suggest that there is a maximum limit to the mass of nuclear DNA so that Bs are tolerated as long as this maximum limit is not exceeded.     

Projecting the effects of climate change on the distribution of maize races and their wild relatives in Mexico

Climate change is expected to be a significant threat to biodiversity, including crop diversity at centers of origin and diversification. As a way to avoid food scarcity in the future, it is important to have a better understanding of the possible impacts of climate change on crops. We evaluated these impacts on maize, one of the most important crops worldwide, and its wild relatives Tripsacum and Teocintes. Maize is the staple crop in Mexico and Mesoamerica, and there are currently about 59 described races in Mexico, which is considered its center of origin . In this study, we modeled the distribution of maize races and its wild relatives in Mexico for the present and for two time periods in the future (2030 and 2050), to identify the potentially most vulnerable taxa and geographic regions in the face of climate change. Bioclimatic distribution of crops has seldom been modeled, probably because social and cultural factors play an important role on crop suitability. Nonetheless, rainfall and temperature still represent a major influence on crop distribution pattern, particularly in rainfed crop systems under traditional agrotechnology. Such is the case of Mexican maize races and consequently, climate change impacts can be expected. Our findings generally show significant reductions of potential distribution areas by 2030 and 2050 in most cases. However, future projections of each race show contrasting responses to climatic scenarios. Several evaluated races show new potential distribution areas in the future, suggesting that proper management may favor diversity conservation. Modeled distributions of Tripsacum species and Teocintes indicate more severe impacts compared with maize races. Our projections lead to in situ and ex situ conservation recommended actions to guarantee the preservation of the genetic diversity of Mexican maize.     

Expression of the sweet protein brazzein in maize for production of a new commercial sweetener

The availability of foods low in sugar content yet high in flavour is critically important to millions of individuals conscious of carbohydrate intake for diabetic or dietetic purposes. Brazzein is a sweet protein occurring naturally in a tropical plant that is impractical to produce economically on a large scale, thus limiting its availability for food products. We report here the use of a maize expression system for the production of this naturally sweet protein. High expression of brazzein was obtained, with accumulation of up to 4% total soluble protein in maize seed. Purified corn brazzein possessed a sweetness intensity of up to 1200 times that of sucrose on a per weight basis. In addition, application tests demonstrated that brazzein-containing maize germ flour could be used directly in food applications, providing product sweetness. These results demonstrate that high-intensity sweet protein engineered into food products can give sweetener attributes useful in the food industry.     

UV-B component of sunlight causes measurable damage in field-grown maize (Zea mays L.): developmental and cellular heterogeneity of damage and repair

Ultraviolet radiation has diverse morphogenetic and damaging effects on plants. The end point of damage is reduced plant growth, but in the short term UV radiation damages specific cellular components. We measured cyclobutane pyrimidine dimers in maize DNA from plants grown in natural solar radiation. Green maize tissues had detectable DNA damage, roots had less damage, and anthers had much more damage than green leaves. This heterogeneity in damage levels may reflect differences in dose received or in damage repair. The architecture of green tissues had no measurable effects on DNA damage levels, as leaf sheath and leaf blade were equivalent. We observed a slight increase in damage levels in plants sampled at the end of the day, but there was no accumulation of damage over the growing season. We measured photoreactivation, and found substantial levels of this light-dependent repair in both the epidermis and inner cell layers of leaves, and in all organelles that contain DNA – the nucleus, chloroplasts and mitochondria. We conclude that maize has efficient mechanisms for photorepair of daily UV-induced DNA damage that prevent accumulation.     

Analysis of genetic diversity of domestic cattle in East and Southeast Asia in terms of variations in restriction sites and sequences of mitochondrial DNA

There are three major groups of domestic cattle in East and Southeast Asia: European cattle, Zebu cattle, and Bali cattle. Ten restriction enzymes were used to analyze restriction site variants in the mitochondrial DNA (mtDNA) in 178 individuals belonging to these three groups of cattle. The results indicate that each of the three groups has mtDNA with a specific haplotype. The sequence of the mitochondrial gene for cytochrome b in representative haplotypes of Zebu and Bali cattle was determined and was compared with that of European cattle in the literature. We calculated 51 pairwise nucleotide sequence differences between European and Zebu cattle and 91 between European and Bali cattle. Our results suggest that ancestral populations of Asiatic domestic cattle may have diverged into two lineages—Bali and European plus Zebu—more than 3 million years ago, and then the European and Zebu groups diverged more than 1 million years or so before domestication occurred.The sequences of mitochondrial genes for cytochrome b from Zebu cattle, Bali cattle and Water buffalo will appear in the DDBJ, EMBL and GenBank nucleotide sequence databases, with the following accession numbers: D34635, D34636 and D34637.