Rapid Evolution of Simple Sequence Repeat Induced by Allopolyploidization

Microsatellite evolution normally occurs in diploids. Until now, there has been a lack of direct experimental evidence for microsatellite evolution following allopolyploidization. In the present study, F₁ hybrids and newly synthesized allopolyploids were derived from Triticum aestivum Chinese Spring × Secale cereale Jinzhou-heimai. One hundred and sixty-three wheat simple sequence repeat (SSR) markers were used to investigate the variation of wheat microsatellites after allopolyploidization and variation of the PCR products of 29 of the SSR markers was observed. Of these 29 SSR markers, 15 were unable to produce products from amphiploids. The other 14 SSR markers did produce products from parental wheat, F₁ hybrids and amphiploids. However, the length of the products amplified from amphiploids was different from the length of the products amplified from parental wheat and F₁ hybrids. Sequencing indicated that the length variation of the 14 microsatellites stemmed mainly from variation in the number of repeat units. The alteration of repeat units occurred in both perfect and compound repeats. In some compound SSR loci, one motif was observed to expand whereas another to contract. Almost all the microsatellite evolution observed in this study could be explained by the slipped-strand mispairing model. The results of this study seem to indicate that stress caused by allopolyploidization might be one of the factors that induce microsatellite evolution. In addition, the findings of present study provided an instance of how simple sequence repeats evolved after allopolyploidization.     

Chromium Improves Glucose Uptake and Metabolism Through Upregulating the mRNA Levels of IR,GLUT4, GS,and UCP3 in Skeletal Muscle Cells

The aim of this study was to evaluate the impact of three different chromium forms as chromic chloride (CrCl), chromium picolinate (CrPic), and a newly synthesized complex of chromium chelated with small peptides (CrSP) on glucose uptake and metabolism in vitro. In cultured skeletal muscle cells, chromium augmented insulin-stimulated glucose uptake and metabolism as assessed by a reduced glucose concentration of culture medium. At the molecular level, insulin significantly increased the mRNA levels of insulin receptor (IR), glucose transporter 4 (GLUT4), glycogen synthase (GS), and uncoupling protein-3 (UCP3), and these impacts can be enhanced by the addition of chromium, especially in the form of CrSP. Collectively, results of this study demonstrate that chromium improves glucose uptake and metabolism through upregulating the mRNA levels of IR, GLUT4, GS, and UCP3 in skeletal muscle cells, and CrSP has higher efficacy on glucose uptake and metabolism compared to the forms of CrCl and CrPic.     

The Decrease of Relative Weight,Lesions, and Apoptosis of Bursa of Fabricius Induced by Excess Dietary Selenium in Chickens

Selenium is an essential trace element possessing immune-stimulatory properties. The purpose of this 42-day study was to investigate the effects of excess dietary sodium selenite on immune function by determining morphological changes and apoptosis of bursa of Fabricius. Three hundred 1-day-old Avian broilers were fed on a basic diet (0.2 ppm selenium) or the same diet amended to contain 1, 5, 10, and 15 ppm selenium supplied as sodium selenite (n = 60/group). Relative weight of bursa was significantly decreased in the 1, 5, 10, and 15 ppm groups at 28 days of age, when compared with that of 0.2 ppm group. Pathological lesions were progressed with the dietary Se level increased. The gross lesions of bursa involved obvious atrophy with decreased volume and pale color. Histopathologically, decreased number of lymphocytes and loosely packed lymphocytes appeared in the medulla and cortex in the follicles. Ultrastructurally, mitochondria injury and increased apoptotic cells with condensed nuclei were observed. In comparison to that of control group, excess Se (5, 10, and 15 ppm) intake increased the percentage of Annexin V positive cells, as measured by flow cytometry. Terminal deoxynucleotidyl transferase 2′-deoxyuridine 5′-triphosphate nick end-labeling assay showed that there were increased frequencies of apoptotic cells in 10 and 15 ppm selenium groups. These data suggest that Se supplementation with sodium selenite should be carefully evaluated as excess selenium (more than 5 ppm) intake could cause profound immunologic inhibition.     

Karyotypic, palynological, and RAPD study on 12 taxa from two subsections of section Idaeobatus in Rubus L. and taxonomic treatment of R. ellipticus, R. pinfaensis, and R. ellipticus var. obcordatus

Twelve taxa belonging to two subsections of section Idaeobatus in Rubus L. from southwestern China were characterized by karyotypic, palynological, and random amplified polymorphic DNA (RAPD) data as follows: (1) The 12 taxa were all diploid species (2n = 2x = 14), among which the chromosome counts for R. mesogaeus var. oxycomus, R. subtibetanus, R. ellipticus var. obcordatus, R. inopertus var. echinocalyx, and R. stans were reported for the first time; (2) All taxa except for R. ellipticus and R. pinfaensis could be distinguished from each other by karyotype, pollen morphology, and RAPD markers. Karyotypes were mainly characterized by the difference in numbers and positions of submetacentric chromosomes and chromosomes with satellited pair, the index of the karyotypic asymmetry, and the ratio of the longest to the shortest chromosome. Pollen morphology were mainly characterized by the discrepancy in specific pollen size, P/E ratio, colpi width, distance between the apices of two ectocolpi, and exine ornamentation characters. The cluster results based on RAPD markers were consistent with morphology classification except for R. pinfaensis; (3) Based on the general data of karyotypic, palynological, and RAPD, R. ellipticus var. obcordatus should be treated as a species R. obcordatus, R. ellipticus and R. pinfaensis should be combined as R. ellipticus, and it was more reasonable to place the combinants and R. obcordatus into subsection Stimulantes rather than into subsection Pungentes.     

Delipidation of insect lipoprotein,lipophorin, affects its binding to the lipophorin receptor,LpR: Implications for the role of LpR-mediated endocytosis

The insect lipophorin receptor (LpR), an LDL receptor (LDLR) homologue that is expressed during restricted periods of insect development, binds and endocytoses high-density lipophorin (HDLp). However, in contrast to LDL, HDLp is not lysosomally degraded, but recycled in a transferrin-like manner, leaving a function of receptor-mediated uptake of HDLp to be uncovered. Since a hallmark of circulatory HDLp is its ability to function as a reusable shuttle that selectively loads and unloads lipids at target tissues without being endocytosed or degraded, circulatory HDLp can exist in several forms with respect to lipid loading. To investigate whether lipid content of the lipoprotein affects binding and subsequent endocytosis by LpR, HDLp was partially delipidated in vitro by incubation with α-cyclodextrin, yielding a particle of buoyant density 1.17 g/mL (HDLp-1.17). Binding experiments demonstrated that LpR bound HDLp-1.17 with a substantially higher affinity than HDLp both in LpR-transfected Chinese hamster ovary (CHO) cells and isolated insect fat body tissue endogenously expressing LpR. Similar to HDLp, HDLp-1.17 was targeted to the endocytic recycling compartment after endocytosis in CHO(LpR) cells. The complex of HDLp-1.17 and LpR appeared to be resistant to endosomal pH, as was recently demonstrated for the LpR–HDLp complex, corroborating that HDLp-1.17 is recycled similar to HDLp. This conclusion was further supported by the observation of a significant decrease with time of HDLp-1.17-containing vesicles after endocytosis of HDLp-1.17 in LpR-expressing insect fat body tissue. Collectively, our results indicate that LpR favors the binding and subsequent endocytosis of HDLp-1.17 over HDLp, suggesting a physiological role for LpR in selective endocytosis of relatively lipid-unloaded HDLp particles, while lipid reloading during their intracellular itinerary might result in decreased affinity for LpR and thus allows recycling.     

毛蕊铁线莲的组织培养与植株再生

1植物名称毛蕊铁线莲（Clematis lasiandra Maxim．）,别名小木通、丝瓜花。2材料类别带芽茎段、节间和叶片。3培养条件诱导培养基：（1）MS＋6-BA0．5mg．L-1（单位下同）＋NAA0．05＋3％蔗糖;（2）MS＋6-BA0．5＋NAA0．1＋2,4-D0．1＋3％蔗糖;（3）MS＋6-BA2．O＋NAA0．1＋3％蔗糖。增殖分化培养基：（4）MS＋6-BA1．0＋NAA0．1＋3％蔗糖;（5）MS＋6．BA2．0＋NAA0．1＋2,4-D0．01＋3％蔗糖;（6）MS＋6．BA2．0＋NAA0．05＋3％蔗糖。生根培养基：（7）1／4MS＋NAA0．5＋0．1％活性炭＋15％蔗糖。所有培养基均附加0．6％琼脂粉,pH5．8-6．0,培养温度为（25±2）℃,光照强度为3040gm01．m-2．S-1,光照时间为14h．d-1。     

Molecular Characterization of the pina Gene in Einkorn Wheat

Fifty-six sequences encoding the pina protein were characterized from three species or subspecies of einkorn wheat. These sequences contained 1,595 nucleotides, including 1,270 conserved sites, 21 single nucleotide polymorphisms (SNPs), and 16 indels. The average frequency of SNPs and indels was one out of 76.1 and 99.9 bases, respectively. Five SNPs and no indels were found in the translated sequences. Fourteen haplotypes were defined, and the accessions in each haplotype ranged from 1 to 18. There were nine haplotypes in Triticum monococcum ssp. aegilopoides, eight in T. monococcum ssp. monococcum, and two in T. urartu. Phylogenetic analysis showed that pina genes from different species or subspecies could be clearly differentiated based on the open reading frame. Genes from T. urartu grouped together, whereas genes from T. monococcum ssp. aegilopoides and T. monococcum ssp. monococcum were shared by three and two clusters, respectively. Both the haplotype and phylogenetic analyses indicated that T. monococcum ssp. aegilopoides was more diverse. These results would contribute to the understanding of functional aspects and efficient utilization of pina genes.     

Genetic Diversity Analysis of Tibetan Wild Barley Using SSR Markers

One hundred and six accessions of wild barley collected from Tibet, China, including 50 entries of the two-rowed wild barley Hordeum vulgare ssp. spontaneum (HS), 29 entries of the six-rowed wild barley Hordeum vulgare ssp. agriocrithon (HA), and 27 entries of the six-rowed wild barley Hordeum vulgare ssp. agriocrithon var. lagunculiforme (HL), were analyzed using 30 SSR markers selected from the seven barley linkage groups for studying genetic diversity and evolutionary relationship of the three subspecies of Tibetan wild barley to cultivated barley in China. Over the 30 genetic loci that were studied, 229 alleles were identified among the 106 accessions, of which 70 were common alleles. H. vulgare ssp. spontaneum possesses about thrice more private alleles (2.83 alleles/locus) than HS (0.93 alleles/locus), whereas almost no private alleles were detected in HL. The genetic diversity among-subspecies is much higher than that within-subspecies. Generally, the genetic diversity among the three subspecies is of the order HS > HL > HA. Phylogenetic analysis of the 106 accessions showed that all the accessions of HS and HA was clustered in their own groups, whereas the 27 accessions of HL were separated into two groups (14 entries with group HS and the rest with group HA). This indicated that HL was an intermediate form between HS and HA. Based on this study and previous works, we suggested that Chinese cultivated barley might evolve from HS via HL to HA.     

Genetic Analysis and Molecular Tagging on a Novel Excessive Tillering Mutant in Rice

A rice (Oryza sativa L.) mutant with an excessive tiller number, designated ext-M1B, was found in the F₂ progenies generated from the cross between M1B and GMS-1 (a genetic male sterile), whose number of tillers was 121. The excessive tillering mutant also resulted in significant changes in plant height, flag leaf, stem, filled grains per panicle, and productive panicles per plant. The inbreeding progenies of ext-M1B exhibited the same mutant phenotype. The crosses from ext-M1B/M1B, M1B/ext-M1B, 2480B/ext-M1B, D62B/ext-M1B, G46B/ext-M1B, and G683B/ext-M1B expressed normal tillering in F₁, and segregated into two different phenotypes of normal tillering type and excessive tillering type in a ratio of 3:1 in F₂. Inheritance analysis indicated that the excessive tillering character was controlled by a single recessive nucleic gene. By BSA (bulked segregants analysis) and microsatellite makers with the F₂ population of 2480B/ext-M1B as the mapping population, RM197, RM584, and RM225, all of which were located on the short arm of rice chromosome 6, were identified to be linked with the excessive tillering gene with genetic distance of 3.8 cM, 5.1 cM, and 5.2 cM, respectively. This gene is probably a new excessive tillering gene in rice and is designated tentatively ext-M1B (t).     

Construction of a Microsatellite Linkage Map with Two Sequenced Rice Varieties

Based on the successful development of new microsatellite markers from the data of two whole-sequenced rice varieties, japonica variety Nipponbare and indica variety 9311, an F₂ population of 90 lines, which was derived from a single cross between Nipponbare and 9311, was applied to construct a genetic linkage framework map. The map covered 2 455.7 cM of total genomic length, and consisted of 152 simple sequence repeats (SSRs) loci including 46 pairs of new SSR primers developed by our research institute. The average genetic distance between two markers was 16.16 cM. In addition, markers RM345 and RM494, which have not been mapped on the Temnykh's map et al. (2001) were anchored on the sixth chromosome of this map. We compared this research with maps of Temnykh et al.(2001) and LAN et al. (2003) regarding the aspects of type and size of population, type and quantity of markers, and the marker arrangement order on chromosome, etc. Results indicated that the similarity of marker linear alignment was 93.81% between this map and T-map, Finally, the important significance of using sequenced rice varieties to construct linkage map was also discussed.