四倍体稗草的组织培养与快速繁殖

以筛选得到的四倍体稗草成熟胚作为外植体,进行了组织培养与快速繁殖技术的研究。结果表明:最佳外植体消毒方式为70%乙醇浸泡30 s+0.1%(w/v)的升汞消毒15 min。最佳诱导培养基为MS盐、DL维生素+2.5 mg/L 2,4-D+300 mg/L谷氨酰胺+500 mg/L脯氨酸+3%麦芽糖。将愈伤组织转入分化培养基(MS+50 mg/L肌醇+1.0 mg/L 6-BA+0.2 mg/L KT+0.5 mg/L NAA+0.25 mg/L IAA+3%麦芽糖)上,培养20 d后超过70%的愈伤组织分化成苗。将3-5 cm长的分化苗转入1/2 MS生根培养基进行生根培养。炼苗后,移入营养土与珍珠岩(2∶1)的基质中,移栽成活率高达90%以上。该体系的建立为稗草抗除草剂基因的功能验证提供了前提条件。     

Isolation and characterization of 17 polymorphic microsatellites in grass carp

Here we report the isolation and characterization of 17 polymorphic loci isolated from a partial genomic DNA library of grass carp (Ctenopharyngodon idellus) enriched for CA repeats. We tested variability of these microsatellites on 24 unrelated individuals collected in China. All microsatellites were polymorphic. The average allele number was 7.9 per locus, ranging from four to 13. The observed heterozygosity was from 0.46 to 0.88 with an average of 0.71, whereas the average expected heterozygosity was 0.78. Sixteen of the 17 microsatellites conformed to Hardy–Weinberg equilibrium, and inherited independently. These microsatellites can be used to study genetic diversity and population structure of wild populations, and facilitate selective breeding of cultured broodstocks.     

Isolation and characterization of microsatellites in Chinese soft‐shelled turtle,Pelodiscus sinensis

Fifteen polymorphic microsatellite loci were developed for the Chinese soft‐shelled turtle (Pelodiscus sinensis) from the (GT)_n microsatellite‐enriched genomic library, using the fast isolation by amplified fragment length polymorphism of sequences containing repeats protocol. The polymorphism of all 15 loci ranged from two to seven alleles with observed heterozygosities ranging from 0.03 to 0.98 (mean 0.43) in one population of 40 individuals. These novel loci will be helpful for understanding the population structure at genetic level and marker‐assisted breeding of this vulnerable species.     

Isolation and characterization of polymorphic microsatellites in a Yangtze River fish,brass gudgeon (Coreius heterodon Bleeker)

The brass gudgeon (Coreius heterodon) is a fish of economic importance in the Yangtze River. From a (GATA)_n‐enriched genomic library, 25 microsatellites were developed by employing the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol. Nine loci exhibited polymorphism with two to 12 alleles (mean 3.9 alleles/locus) in a test population, and observed heterozygosity ranged from 0.1111 to 0.9630 (mean 0.4426). Three of the nine loci showed polymorphism in a congeneric species, the largemouth bronze gudgeon Coreius guichenoti. These loci should provide sufficient level of genetic diversity to evaluate the fine‐scale population structure of C. heterodon.     

Seed defensins of barnyard grass Echinochloa crusgalli (L.) Beauv

From the annual weed barnyard grass Echinochloa crusgalli (L.) Beauv., two novel defensins Ec-AMP-D1 and Ec-AMP-D2 that differ by a single amino acid substitution were isolated by a combination of different chromatographic procedures. Both defensins were active against several phytopathogenic fungi and the oomycete Phytophthora infestans at micromolar concentrations. The Ec-AMP-D1 showed higher activity against the oomycete than Ec-AMP-D2. The comparison of the amino acid sequences of the antifungal E. crusgalli defensins with those of earlier characterized T. kiharae defensins [T.I. Odintsova, Ts.A. Egorov, A.Kh. Musolyamov, M.S. Odintsova, V.A. Pukhalsky, E.V. Grishin, Seed defensins from T. kiharae and related species: genome localization of defensin-encoding genes, Biochimie, 89 (2007) 605-612.] that were devoid of substantial antifungal activity point to the C-terminal region of the molecule as the main determinant of the antifungal activity of E. crusgalli defensins.     

Isolation and characterization of microsatellites in the Southeast Asian river catfish Mystus nemurus

A total of 26 simple sequence repeats were identified using a random amplified polymorphic DNA (RAPD) based technique in the Southeast Asian river catfish Mystus nemurus. We report on the characterization of five polymorphic microsatellite loci in M. nemurus. The average number of alleles per locus was 3.2. These are the first microsatellite loci that have been developed for this species. These markers should prove useful as tools for managing the brood stocks and for future aquacultural development of this species.     

Isolation and characterization of trinucleotide microsatellites in African malaria mosquito Anopheles funestus

We developed microsatellite markers for an important African malaria mosquito Anopheles funestus Giles. The microsatellite‐enriched genomic library was constructed and screened with single‐strand oligonucleotides [(CCT)₁₇, (AAT)₁₇, (CAG)₁₇ and (GA)₂₅] as probes. Among the 47 pairs of polymerase chain reaction primers screened, 31 produced successful and consistent amplification. Although only a few A. funestus individuals from one geographical location were used to screen microsatellite marker polymorphism, 27 markers were found polymorphic and four markers monomorphic. Most polymorphic markers are trinucleotide markers. Isolation of polymorphic microsatellite markers provide useful tools for A. funestus population genetic studies and genome mapping.     

Isolation and characterization of polymorphic microsatellites in a sex‐reversal fish,rice field eel (Monopterus albus)

The rice field eel (Monopterus albus) is a fish of economic importance in China and some Asian countries. From a (GT)_n‐enriched genomic library, 30 microsatellites were developed by employing the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol. Thirteen loci exhibited polymorphism with two to 13 alleles (mean 7.9 alleles/locus) in a test population and observed heterozygosity ranged from 0.3125 to 0.9688 (mean 0.7140). These loci should provide sufficient level of genetic variation to study the fine‐scale population structure and reproductive ecology of the species.     

Isolation and characterization of microsatellites from the canine genome

Microsatellite sequences comprising (dC-dA)_n.(dG-dT)_n repeats have been isolated from canine libraries and sequenced. Oligonucleotide primers have been synthesized to the micro-satellite flanking sequences and used in the polymerase chain reaction to amplify those loci from genomic DNA. The degree of polymorphism of each microsatellite was estimated in a set of unrelated dogs. It is concluded that of the 10 loci studied, nine are sufficiently polymorphic to be useful in genetic studies.     

Isolation and characterization of microsatellite markers in the sacred lotus (Nelumbo nucifera Gaertn.)

The sacred lotus (Nelumbo nucifera Gaertn.) is an aquatic plant of economic and ornamental importance in China. From an (AG)_n‐enriched genomic library, 24 microsatellites were isolated and identified by using the (fast isolation by the AFLP of sequences containing repeats) FIASCO protocol. Eleven loci showed polymorphism with two to six alleles per locus. These markers yielded 42 alleles in a survey of 32 accessions of the sacred lotus. Eleven effective primer pairs of simple sequence repeats were designed and will be used as genetic markers to evaluate the fine‐scale population structure of the sacred lotus in the future.     

Isolation and characterization of microsatellites in Zosterisessor ophiocephalus (Perciformes,Gobiidae)

We isolated and characterized 10 polymorphic microsatellite loci from two partial genomic libraries of Zosterisessor ophiocephalus (Perciformes, Gobiidae), enriched for AC and for multiple motifs. Variability was tested on specimens from the Venice lagoon revealing a high degree of genetic variation. The loci will be useful for monitoring the biodiversity of the species at the genetic level and for parentage assessment purposes.     

Isolation and characterization of sequence‐tagged microsatellite sites markers in chickpea (Cicer arietinum L.)

In this study we report the isolation of microsatellite sequences and their conversion to sequence‐tagged microsatellite sites (STMS) markers in chickpea (Cicer arietinum L.). Thirteen putative recombinants isolated from a chickpea genomic library were sequenced, and used to design 10 STMS primer pairs. These were utilized to analyse the genetic polymorphism in 15 C. arietinum varieties and two wild varieties, C. echinospermum and C. reticulatum. All the primer pairs amplified polymorphic loci ranging from four to seven alleles per locus. The observed heterozygosity ranged from 0 to 0.6667. Most of the STMS markers also amplified corresponding loci in the wild relatives suggesting conservation of these markers in the genus. Hence, these polymorphic markers will be useful for the evaluation of genetic diversity and molecular mapping in chickpea.     

稗类(Echinochloa spp.)植物的开花生物学特性

对稗类（Echinochloa spp.）常见种类[稗Echinochloa crusgalli(L.)Beauv.;无芒稗 E.crusgalli var.mitis(Pursh)Peterm.;西来稗E.crusgalli var.zelayensis(H.B.K.)Hitchc.;细叶旱稗E.crusgalli var.praticola Ohwi;旱稗E.hispidula(Retz.)Nees和光头稗E.colonum(L.)Link]的开花过程和节律,温度、光周期、晴天和阴雨天对其中部分稗类植物开花的影响以及套袋条件下无芒稗和旱稗的结实率进行了研究。结果表明,不同稗类植物的开花的过程基本相似,从颖壳微微张口到闭合大约需20－30min,但花约伸出颖壳后,散落花粉的过程很快,颖壳闭合后柱头外露率达100％。稗类植物每穗开花持续时间较长,一般为20－25d,随种类的不同略有变化;稗类植物每穗每天开花数量的变化趋势基本呈抛物线状,每天的开花比例变化较平缓,即使在开花高峰期,开花数也只占总数的10％－15％,开花高峰期因稗草种类的不同略有变化,但都在始花后11－15d左右。每日的开花节律为70％－80％的颖花在7：00之前开,15％－20％在7：00－8：30开,8：30之后零星开花,10：30尚未见开花。温度、光周期、晴天和阴雨天对开花都有一定的影响。不同条件下的结实率没有明显差异。     

Thermotropic Properties of Cellular Membranes in Dormant and Non-Dormant Echinochloa crus-galli(L.) Beauv. Seeds

Steady-state fluorescence polarization measurements with l,6-diphenyl-l,3,5-hexatriene(DPH) were used to monitor thermotropic transitions in microsomalfractions and plasma membrane vesicles isolated from barnyardgrass[Echinochloa crus-galli (L.) Beauv.] seeds during the transititionfrom dormancy to germination. The effect of dormancy-relievingor inactive alcohols on the thermotropic properties of the cellularmembranes was determined both in vivo and in vitro. Membranefractions isolated from dormant seeds showed some discontinuitiesin the Arrhenius plots. In non-dormant or germinating seedscellular membranes showed linear Arrhenius plots over the entirerange of temperature examined. Membrane preparations from imbibedseeds showed a similar pattern in their Arrhenius plots upontreatment with the various alcohols in vitro. The results suggestthat the release from dormancy in seeds is associated with somechanges in their cellular membranes. Key words: Germination, alcohols, thermotropic transition     

Isolation and characterization of microsatellites in the bryozoan Crisia denticulata

Cyclostomata bryozoa are thought to reproduce via polyembryony, a clonal replication of a fertilized egg. To test this hypothesis and to assess the impact of their reproductive strategy on the structure of populations, we isolated microsatellite markers in Crisia denticulata (Cyclostomata, Stenolemata), using an initial enrichment step with random amplified polymorphic DNA (RAPD) primers. A total of nine microsatellites, one tetra‐ and eight dinucleotides repeats were isolated; seven were found to be polymorphic in a test sample of 30 individuals, with allele numbers/locus varying from 2 to 6. The tetranucleotide locus showed heterozygote deficiency. These primers did not amplify the DNA of Crisia eburnea.     

Isolation and characterization of microsatellites in the Adriatic sturgeon (Acipenser naccarii)

We isolated and characterized seven polymorphic microsatellite loci from a partial genomic library of the Adriatic sturgeon (Acipenser naccarii) enriched for GATA repeats. Variability was tested in 20 specimens of this endangered species. Our data support the tetraploid condition for A. naccarii and will be useful for the investigation of the remnant genetic variability of this species as well as for genetic tagging studies.