SRAP标记与形态学标记在西瓜DUS测试中的比较

DUS（特异性、一致性和稳定性）测试是进行新品种申请的必要步骤。本文以28个不同的西瓜品种为研究对象,分别采用21对SRAP引物标记和54个用于DUS测试的形态学标记对其进行遗传多样性分析,其中SRAP引物在不同品种间的多态信息含量（PIC）在52.5%~89.2%之间,平均值为72.0%,计算得到的各材料间相似系数在0.92至0.99之间,而形态学标记统计得到各材料间相关系数在0.50到0.85之间。采用UPGMA法对所有材料进行聚类分析,SRAP分子标记聚类划分成四类,形态学标记将其划分为五类。对两种标记所得的结果进行相关性分析得出两者的相关系数为0.218,表明形态学标记和SRAP标记在这些材料上表现的相关性不是很高,但在品种鉴定和区别上SRAP标记表现出一定的优势,可以作为DUS测试的一种有益补充。     

Ethylene biosynthetic genes are differentially expressed during carnation (Dianthus caryophyllus L.) flower senescence

Ethylene production and expression patterns of an 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase (CARAO1) and of two ACC synthase (EC 4.4.1.14) genes (CARACC3 and CARAS1) were studied in floral organs of cut carnation flowers (Dianthus caryophyllus L.) cv. White Sim. During the vase life and after treatment of fresh flowers with ethylene, production of ethylene and expression of ethylene biosynthetic genes first started in the ovary followed by the styles and the petals. ACC oxidase was expressed in all the floral organs whereas, during the vase life, tissue-specific expression of the two ACC synthase genes was observed. After treatment with a high ethylene concentration, tissue specificity of the two ACC synthase genes was lost and only a temporal difference in expression remained. In styles, poor correlation between ethylene production and ACC synthase (CARAS1) gene expression was observed suggesting that either activity is regulated at the translational level or that the CARAS1 gene product requires an additional factor for activity.Isolated petals showed no increase in ethylene production and expression of ethylene biosynthetic genes when excised from the flower before the increase in petal ethylene production (before day 7); showed rapid cessation of ethylene production and gene expression when excised during the early phase of petal ethylene production (day 7) and showed a pattern of ethylene production and gene expression similar to the pattern observed in the attached petals when isolated at day 8. The interorgan regulation of gene expression and ethylene as a signal molecule in flower senescence are discussed.     

Divergent fructokinase genes are differentially expressed in tomato.

Two cDNA clones (Frk1 and Frk2) encoding fructokinase (EC 2.7.1.4) were isolated from tomato (Lycopersicon esculentum). The Frk2 cDNA encoded a deduced protein of 328 amino acids that was more than 90% identical with a previously characterized potato (Solanum tuberosum) fructokinase. In contrast, the Frk1 cDNA encoded a deduced protein of 347 amino acids that shared only 55% amino acid identity with Frk2. Both deduced proteins possessed and ATP-binding motif and putative substrate recognition site sequences identified in bacterial fructokinases. The Frk1 cDNA was expressed in a mutant yeast (Saccharomyces cerevisiae) line, which lacks the ability to phosphorylate glucose and fructose and is unable to grow on glucose or fructose. Mutant cells expressing Frk1 were complemented to grow on fructose but not glucose, indicating that Frk1 phosphorylates fructose but not glucose, and this activity was verified in extracts of transformed yeast. The mRNA corresponding to Frk2 accumulated to high levels in young, developing tomato fruit, whereas the Frk1 mRNA accumulated to higher levels late in fruit development. The results indicate that fructokinase in tomato is encoded by two divergent genes, which exhibit a differential pattern of expression during fruit development.     

Two tomato non-symbiotic haemoglobin genes are differentially expressed in response to diverse changes in mineral nutrient status

The role of plant non‐symbiotic haemoglobins remains undefined, but recent findings suggest a possible role in plant nitrate nutrition. This study sought to characterize patterns of gene expression for two tomato (Lycopersicon esculentum L.) non‐symbiotic haemoglobin genes, and to examine how diverse changes in nutrient status influences the expression of these two genes. The deduced amino acid sequences of the genes examined, SOLly GLB1 and SOLly GLB2, are 55% identical. SOLly GLB1 mRNA is present in highest abundance in roots and older stems, whereas SOLly GLB2 mRNA abundance is highest in leaves, but detectable in other tissues including roots. The SOLly GLB1 mRNA levels increased rapidly in roots in response to each nutrient treatment examined in hydroponic culture, including the individual removal of phosphate, potassium and iron from the culture medium, as well as by the addition of nitrate. In contrast, the levels of SOLly GLB2 mRNA were not significantly altered in response to these same treatments. These results are the first report that non‐symbiotic haemoglobin gene expression can be influenced by a broad range of changes both in mineral macronutrient and micronutrient status, suggesting a previously unrecognized generic role for non‐symbiotic haemoglobins in processes associated with mineral nutrient nutrition.     

西瓜种质资源遗传亲缘关系的RAPD分析

利用 RAPD 技术对国内外32份西瓜主栽品种与其骨干亲本及野生类型的遗传亲缘关系进行了研究。从720个随机引物(10bp)中筛选出15个能产生稳定多态性的引物用于 RAPD 反应,共扩增出104条 DNA 带,其中多态性 DNA 条带43条,占41.35%,平均每个引物扩增的 DNA 条带的数目为7.0条。聚类分析将供试材料分为6个类群:1个东亚生态型类群、1个美国生态型类群、2个中间生态型类群和2个非洲野生型类群,与传统的西瓜生态型分类基本吻合。每个生态型类群都有其特有的扩增(缺失)条带,同时分析了同一生态型中各个品种之间的亲缘关系及其品种的特异条带。本实验结果不仅从分子水平验证了西瓜是遗传基础狭窄的作物,而且在分子水平对西瓜传统分类与地理生态型分类进行了分析。     

Tumor promoter-inducible genes are differentially expressed in the developing mouse.

TIS genes are rapidly and transiently induced by tetradecanoyl phorbol acetate in 3T3 cells. We analyzed the developmental appearance of a number of the TIS genes to determine whether, in a normal physiological context, these genes have common or distinct mechanisms of regulation. Each TIS gene has a distinct tissue specificity and/or developmental profile.     

Two isoforms of the GBSSI class of granule-bound starch synthase are differentially expressed in the pea plant (Pisum sativum L.)

In addition to the GBSSI isoform of starch synthase described previously, the pea plant contains a second, granule-bound isoform, GBSSIb. GBSSI is abundant in pea embryos and Rhizobium root nodules, is present at low levels in pods and is absent from leaves. Mutations at the lam locus eliminate GBSSI from all of these organs. GBSSIb is present in pods, leaves and nodules and is unaffected by mutations at the lam locus. GBSSI and GBSSIb are very similar in molecular mass, primary sequence, activity and antigenic properties. GBSSIb, like GBSSI, can synthesize amylose in the presence of malto-oligosaccharides in isolated starch granules. However, its role in vivo is unclear. The lam mutation eliminates amylose from the starch of embryos but does not affect the relatively small amounts of amylose-like material in the starch of pods, leaves and nodules. The significance of these results for understanding of the regulation of amylose synthesis is discussed.     

Identification of genes differentially expressed in hemocytes of Scylla paramamosain in response to lipopolysaccharide

Although the crab Scylla paramamosain has been cultured in China for a long time, little knowledge is available on how crabs respond to infection by bacteria. A forward suppression subtractive hybridization (SSH) cDNA library was constructed from their hemocytes and the up-regulated genes were identified in order to isolate differentially expressed genes in S. paramamosain in response to bacterial lipopolysaccharide (LPS). A total of 721 clones on the middle scale in the SSH library were sequenced. Among these genes, 271 potentially functional genes were recognized based on the BLAST searches in NCBI and were categorized into seven groups in association with different biological processes using AmiGO against the Gene Ontology database. Of the 271 genes, 269 translatable DNA sequences were predicted to be proteins, and the putative amino acid sequences were searched for conserved domains and proteins using the CD-Search service and BLASTp. Among 271 genes, 179 (66.1%) were annotated to be involved in different biological processes, while 92 genes (33.9%) were classified as an unknown-function gene group. It was noted that only 18 of the 271 genes (6.6%) had previously been reported in other crustaceans and most of the screened genes showed less similarity to known sequences based on BLASTn results, suggesting that 253 genes were found for the first time in S. paramamosain. Furthermore, two up-regulated genes screened from the SSH library were selected for full-length cDNA sequence cloning and in vivo expression study, including Sp-superoxide dismutase (Sp-Cu-ZnSOD) gene and Sp-serpin gene. The differential expression pattern of the two genes during the time course of LPS challenge was analyzed using real-time PCR. We found that both genes were significantly expressed in LPS-challenged crabs in comparison with control. Taken together, the study primarily provides the data of the up-regulated genes associated with different biological processes in S. paramamosain in response to LPS, by which the interesting genes or proteins potentially involved in the innate immune defense of S. paramamosain will be investigated in future.     

Identification of differentially expressed genes in response to dietary iron deprivation in rat duodenum

We sought to identify novel genes involved in intestinal iron absorption by inducing iron deficiency in rats during postnatal development from the suckling period through adulthood. We then performed comparative gene chip analyses (RAE230A and RAE230B chips; Affymetrix) with cRNA derived from duodenal mucosa. Real-time PCR was used to confirm changes in gene expression. Genes encoding the apical iron transport-related proteins [divalent metal transporter 1 (DMT1) and duodenal cytochrome b] were strongly induced at all ages studied, whereas increases in mRNA encoding the basolateral proteins iron-regulated gene 1 and hephaestin were observed only by real-time PCR. In addition, transferrin receptor 1 and heme oxygenase 1 were induced. We also identified induction of novel genes not previously associated with intestinal iron transport. The Menkes copper ATPase (ATP7a) and metallothionein were strongly induced at all ages studied, suggesting increased copper absorption by enterocytes during iron deficiency. We also found significantly increased liver copper levels in 7- to 12-wk-old iron-deficient rats. Also upregulated at most ages examined were the sodium-dependent vitamin C transporter, tripartite motif protein 27, aquaporin 4, lipocalin-interacting membrane receptor, and the breast cancer-resistance protein (ABCG2). Some genes also showed decreased expression with iron deprivation, including several membrane transporters, metabolic enzymes, and genes involved in the oxidative stress response. We speculate that dietary iron deprivation leads to increased intestinal copper absorption via DMT1 on the brush-border membrane and the Menkes copper ATPase on the basolateral membrane. These findings may thus explain copper loading in the iron-deficient state. We also demonstrate that many other novel genes may be differentially regulated in the setting of iron deprivation.     

Identification of differentially expressed genes in Con A-activated carp (Cyprinus carpio L.) leucocytes

A cDNA library was constructed from the message RNA (mRNA) obtained from Con A-induced head kidney (HK) leucocytes of carp (Cyprinus carpio L.). Differential screening of the cDNA was carried out by hybridization against the total cDNA probes from normal, Con A-uninduced HK leucocytes or Con A-induced HK leucocytes of carp. The differential expression patterns of certain cDNA clones were confirmed by Southern-blot and Northern-blot analysis. Single-pass of the sequencing analysis and homology search in Genbank (EMBL) revealed those differentially expressed cDNA clones encode for cytochrome c oxidase sub-unit II and III (COII and COIII), elongation factor-1 beta (EF-1 beta), bleomycin hydrolase (BH), heat shock cognate protein 70 (HSC70) and 16S ribosomal RNA (16S rRNA).     

Two cinnamoyl-CoA reductase (CCR) genes from Arabidopsis thaliana are differentially expressed during development and in response to infection with pathogenic bacteria.

Cinnamoyl-CoA reductase (CCR; EC 1.2.1.44) catalyses the conversion of cinnamoyl-CoAs into their corresponding cinnamaldehydes, i.e. the first step of the phenylpropanoid pathway specifically dedicated to the monolignol biosynthetic branch. In previous work, we described the isolation and characterisation of the first cDNA encoding CCR in Eucalyptus (Lacombe, E., Hawkins, S., Van Dorsselaere, J., Piquemal, J., Goffner, D., Poeydomenge, O., Boudet, A.M., Grima-Pettenati, J., 1997. Cinnamoyl CoA reductase, the first committed enzyme of the lignin branch biosynthetic pathway: cloning, expression and phylogenetic relationships. Plant Journal 11, 429--441) and shown the role of this enzyme in controlling the carbon flux into lignins (Piquemal, J., Lapierre, C., Myton, K., O'Connell, A., Schuch, W., Grima-Pettenati, J., Boudet, A.M., 1998. Down-regulation of cinnamoyl-CoA reductase induces significant changes of lignin profiles in transgenic tobacco plants. Plant Journal 13, 71--83). Here, we report the characterisation of two functionally and structurally distinct cDNA clones, AtCCR1 and AtCCR2 (81.6% protein sequence identity) in Arabidopsis thaliana. The two recombinant proteins expressed in Escherichia coli are able to use the three cinnamoyl-CoAs tested but with different levels of efficiency. AtCCR1 is five times more efficient with feruloyl-CoA and sinapoyl-CoA than AtCCR2. In addition, the two genes are differentially expressed during development and in response to infection. AtCCR1 is preferentially expressed in tissues undergoing lignification. In contrast, AtCCR2, which is poorly expressed during development, is strongly and transiently induced during the incompatible interaction with Xanthomonas campestris pv. campestris leading to a hypersensitive response. Altogether, these data suggest that AtCCR1 is involved in constitutive lignification whereas AtCCR2 is involved in the biosynthesis of phenolics whose accumulation may lead to resistance.     

Chlamydomonas reinhardtii has a small family of purple acid phosphatase homologue genes that are differentially expressed in response to phytate

Purple acid phosphatases (PAPs) are metallophosphoesterase enzymes involved in the acquisition and recycling of phosphorus. PAP phytases from microorganisms and plants are responsible for the dephosphorylation of phytate. Phytate is the main storage form of phosphorus in plant seeds and constitutes the major form of organic phosphorus present in soil. Although some phosphatases have been studied in Chlamydomonas reinhardtii, no gene coding for PAPs have so far been characterized. In this study, six PAP homologue genes were identified and characterized in silico in C. reinhardtii (CrPAP1 to CrPAP6). A metallophosphoesterase domain including the seven conserved residues characteristic of PAP enzymes was found in all six CrPAPs. The phylogenetic tree comprising PAP homologue sequences from microalgae, plants, and animals showed nine major clades and CrPAPs resolved in four of them. A constitutive expression was found for CrPAP2, CrPAP3, CrPAP4, and CrPAP6 in all media tested, while CrPAP1 and CrPAP5 were induced by the addition of phytate in a medium without phosphate salts. Our results provide a starting point for further functional analysis of the CrPAP gene family, and the evaluation of their potential as phytases in C. reinhardtii.