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Now showing items 1 - 7 of 7

  • Determination of genetic diversity among Saccharina germplasm using ISSR and RAPD markers

    Cui, Cuiju   Li, Yan   Liu, Yanling   Li, Xiaojie   Luo, Shiju   Zhang, Zhuangzhi   Wu, Ruina   Liang, Guangjin   Sun, Juan   Peng, Jie   Tian, Pingping  

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  • Stable chloroplast transformation of immature scutella and inflorescences in wheat (Triticum aestivum L.) (Retracted article. See vol. 44, pg. 373, 2012)

    Cui, Cuiju   Song, Fei   Tan, Yi   Zhou, Xuan   Zhao, Wen   Ma, Fengyun   Liu, Yunyi   Hussain, Javeed   Wang, Yuesheng   Yang, Guangxiao  

    Chloroplast transformation in wheat was achieved by bombardment of scutella from immature embryos and immature inflorescences, respectively. A wheat chloroplast site-specific expression vector, pBAGNRK, was constructed by placing an expression cassette containing neomycin phosphotransferase II (nptII) and green fluorescent protein (gfp) as selection and reporter genes, respectively, in the intergenic spacer between atpB and rbcL of wheat chloroplast genome. Integration of gfp gene in the plastome was identified by polymerase chain reaction (PCR) analysis and Southern blotting using gfp gene as a probe. Expression of GFP protein was examined by western blot. Three positive transformants were obtained and the Southern blot of partial fragment of atpB and rbcL (targeting site) probes verified that one of them was homoplasmic. Stable expression of GFP fluorescence was confirmed by confocal microscopy in the leaf tissues from T-1 progeny seedlings. PCR analysis of gfp gene also confirmed the inheritance of transgene in the T-1 progeny. These results strengthen the feasibility of wheat chloroplast transformation and also give a novel method for the introduction of important agronomic traits in wheat through chloroplast transformation.
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  • A genome screen for the development of sex-specific DNA markers in Saccharina japonica

    Zhang, Linan   Cui, Cuiju   Li, Yan   Wu, Hao   Li, Xiaojie  

    Saccharina japonica is an important cultured marine brown alga and it has been the subject of intensive genetic improvement during its domestication and farming. However, there are limited credible and effective sex markers to identify the sex of gametophytes and sporophytes derived from normal hybridization or monogenesis. In the present study, the transcriptomic and genomic sequences of S. japonica were screened by 15 sex-determining region (SDR) genes from a model brown alga (Ectocarpus), and six cDNA sequences and their corresponding genomic sequences were identified. Thirteen primer pairs were designed from the coding region of six genomic sequences and subsequently tested on gametophytes of S. japonica. From these regions, four novel male-specific markers were developed, which derived from S. japonica genomic sequences JXRI01001736 and JXRI01002827, corresponding to a male-specific SDR gene and a gametologue SDR gene in Ectocarpus, respectively. In addition, an existing female marker (M_68_58_2) for Macrocystis pyrifera and Undaria pinnatifida, derived from the Ectocarpus SDR gene, could also be used as a female-specific marker for S. japonica. The results of the present study not only demonstrate an important new tool to determine the sex of gametophytes and distinguish between diploid sporophytes and partheno-sporophytes in breeding programmes but also provide insight into the sex-determination system for S. japonica, including the identification of sex chromosomes, SDR, and SDR genes and evolution of sex chromosomes.
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  • Breeding of an intraspecific kelp hybrid Dongfang no. 6 (Saccharina japonica,Phaeophyceae,Laminariales) for suitable processing products and evaluation of its culture performance

    Li, Xiaojie   Zhang, Zhuangzhi   Qu, Shancun   Liang, Guangjin   Zhao, Nan   Sun, Juan   Song, Shaofeng   Cao, Zengmei   Li, Xia   Pan, Jinhua   Luo, Shiju   Zhang, Linan   Cui, Cuiju   Peng, Jie   Li, Yan   Wu, Ruina   Zhao, Juping   Qian, Rui   Wang, Liqin   Sai, Shan   Yang, Guanpin  

    Dongfang no. 6, an intraspecific hybrid of kelp (Saccharina japonica) which was high in yield and had a high stress tolerance and was suitable for processing, was bred by crossing scientifically selected parental gametophyte clones. The sporophyte of the hybrid was raised by crossing a female gametophyte clone isolated from a Korean ecotype of S. japonica (Korean kelp) with a male gametophyte clone isolated from Lianza no. 1, a variety of S. japonica bred through continuous selection. The culture performance, stress tolerance, and suitability for processing of the hybrid and two commercial controls were compared for a period lasting 5 years (2009-2013). It was found that the hybrid was fast in growth, tolerant to high seawater temperature, resistant to strong solar irradiation, late in maturing time, and high in processed product yield. The percentage of salting processed product of the hybrid was 17.5 and 15.4 % higher than those of control 1 and 2, respectively. The yield of salting processed product of the hybrid increased by 24.2 and 38.8 % over control 1 and 2, respectively. The yield of air drying processed product of the hybrid increased by 109.4 and 88.1 % over control 1 and 2, respectively. The breeding of Dongfang no. 6 demonstrated that crossing gametophyte clones in combination with selecting desirable hybrids are highly effective for breeding kelp hybrids with high-culture performance and suitability for processing.
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  • A census of nuclear cyanobacterial recruits in the plant kingdom.

    Makai, Szabolcs   Li, Xiao   Hussain, Javeed   Cui, Cuiju   Wang, Yuesheng   Chen, Mingjie   Yang, Zhaowan   Ma, Chuang   Guo, An-Yuan   Zhou, Yanhong   Chang, Junli   Yang, Guangxiao   He, Guangyuan  

    The plastids and mitochondria of the eukaryotic cell are of endosymbiotic origin. These events occurred ~2 billion years ago and produced significant changes in the genomes of the host and the endosymbiont. Previous studies demonstrated that the invasion of land affected plastids and mitochondria differently and that the paths of mitochondrial integration differed between animals and plants. Other studies examined the reasons why a set of proteins remained encoded in the organelles and were not transferred to the nuclear genome. However, our understanding of the functional relations of the transferred genes is insufficient. In this paper, we report a high-throughput phylogenetic analysis to identify genes of cyanobacterial origin for plants of different levels of complexity: Arabidopsis thaliana, Chlamydomonas reinhardtii, Physcomitrella patens, Populus trichocarpa, Selaginella moellendorffii, Sorghum bicolor, Oryza sativa, and Ostreococcus tauri. Thus, a census of cyanobacterial gene recruits and a study of their function are presented to better understand the functional aspects of plastid symbiogenesis. From algae to angiosperms, the GO terms demonstrated a gradual expansion over functionally related genes in the nuclear genome, beginning with genes related to thylakoids and photosynthesis, followed by genes involved in metabolism, and finally with regulation-related genes, primarily in angiosperms. The results demonstrate that DNA is supplied to the nuclear genome on a permanent basis with no regard to function, and only what is needed is kept, which thereby expands on the GO space along the related genes. =20
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  • Development of genic SSR markers from an assembled Saccharina japonica genome

    Peng, Jie   Zhang, Linan   Li, Xiaojie   Cui, Cuiju   Wu, Ruina   Tian, Pingping   Li, Yan   Liu, Yanling  

    Saccharina japonica, a marine brown alga, is an economically important species that has been cultivated in China for approximately a century. We identified 11,973 simple sequence repeats (SSRs) in 58.3 Mb of predicted coding sequence from an assembled S. japonica genome using Illumina paired-end sequencing data. Trinucleotide SSRs were the most abundant motif. Twenty-six loci amplified by the 57 unique primer pairs we designed from trinucleotide genic SSRs (repeat number > 10) were polymorphic, among the 55 S. japonica individuals tested. The number of alleles per locus ranged from 2 to 7 (average 3.46). The observed and expected heterozygosity per locus ranged from 0.128 to 0.652 and from 0.130 to 0.676, respectively. Two loci deviated from the Hardy-Weinberg equilibrium and two others were in linkage disequilibrium. We demonstrate that genic SSRs can be efficiently identified from assembled and annotated genomes using Illumina sequencing data. The novel polymorphism markers that we have identified, with informative flanking sequences and unique positional relationships to the genome, should facilitate genetic diversity analysis and further genetic studies in this species.
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  • Determination of genetic diversity among Saccharina germplasm using ISSR and RAPD markers

    Cui, Cuiju   Li, Yan   Liu, Yanling   Li, Xiaojie   Luo, Shiju   Zhang, Zhuangzhi   Wu, Ruina   Liang, Guangjin   Sun, Juan   Peng, Jie   Tian, Pingping  

    Various species of genus Saccharina are economically important brown macroalgae cultivated in China. The genetic background of the conserved Saccharina germplasm was not clear. In this report, DNA-based molecular markers such as inter simple sequence repeats (ISSR) and random amplified polymorphic DNA (RAPD) were used to assess the genetic diversity and phylogenetic relationships among 48 Saccharina germplasms. A total of 50 ISSR and 50 RAPD primers were tested, of which only 33 polymorphic primers (17 ISSR and 16 RAPD) had an amplified clear and reproducible profile, and could be used. Seventeen ISSR primers yielded a total of 262 bands, of which 256 were polymorphic, and 15.06 polymorphic bands per primer were amplified from 48 kelp gametophytes. Sixteen RAPD primers produced 355 bands, of which 352 were polymorphic, and 22 polymorphic bands per primer were observed across 48 individuals. The simple matching coefficient of ISSR, RAPD and pooled ISSR and RAPD dendrograms ranged from 0.568 to 0.885, 0.670 to 0.873, and 0.667 to 0.862, revealing high genetic diversity. Based on the unweighted pair group method with the arithmetic averaging algorithm (UPGMA) cluster analysis and the principal components analysis (PCA) of ISSR data, the 48 gametophytes were divided into three main groups. The Mantel test revealed a similar polymorphism distribution pattern between ISSR and RAPD markers, the correlation coefficient r was 0.62, and the results indicated that both ISSR and RAPD markers were effective to assess the selected gametophytes, while matrix correlation of the ISSR marker system (r =3D 0.78) was better than that of the RAPD marker system (r =3D 0.64). Genetic analysis data from this study were helpful in understanding the genetic relationships among the selected 17 kelp varieties (or lines) and provided guidance for molecular-assisted selection for parental gametophytes of hybrid kelp breeding. (C) 2016 Academie des sciences. Published by Elsevier Masson SAS. All rights reserved.
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