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1.
mRNA 差异显示技术是分析差异表达基因的强有力的工具。本实验针对油菜差别显示体系筛选优化,最终获得了差别显示体系。研究结果为：RNA模板在2μg,反转录产物用量为1.5μl时,可以扩增出较多且清晰的条带;PCR退火温度在40℃扩增条带清晰。该体系用于甘蓝型油菜低磷胁迫前后基因差异表达研究稳定可靠。 相似文献
2.
Poncirus trifoliata, a close relative of Citrus species, has been widely used as a cold hardy rootstock for citrus production in low-temperature
environments. cDNA Amplified Fragment Length Polymorphism (cDNA-AFLP) and Relative Quantification (RQ-PCR) were used to study
gene expression of P. trifoliata in response to 10, 24 and 55 h of low temperature (4°C) treatment and a gradual 3 weeks long cold-acclimation regime. Thirteen
differentially expressed cDNA fragments were chosen for further study. These fragments show high similarities to genes with
the following known functions: signal transduction and regulation of gene expression, osmotic stress response, cell skeleton
reorganization, vesicle trafficking and senescence. Expression levels of genes implicated in signal transduction and regulation
of gene expression (MAPK3 and bZIP TF) increased with prolonged exposure to low temperature, but were not upregulated following
cold-acclimation. Others were induced following cold-acclimation, but not upon exposure to 4°C. Only the auxin-ethylene GH3-like
protein showed dramatic increases during all cold treatments. These comparisons provide information on the dynamic changes
in gene expression levels during different cold response regimes. 相似文献
3.
Gregory B. Martin 《Euphytica》1994,79(3):187-193
Tomato (Lycopersicon esculentum) cultivars that contain the Pto bacterial resistance locus also exhibit sensitivity to fenthion, an organophosphorous insecticide. Resistance to Pseudomonas syringae pv. tomato (Pst) encoded by the Pto gene and sensitivity to fenthion cosegregate in large F2 populations and were apparently introgressed together into tomato from the wild species Lycopersion pimpinellifolium. In order to isolate the genes responsible for these two phenotypes and to study their molecular basis, a multistep map-based cloning project was initiated. A closely linked RFLP marker was used to isolate a yeast artificial chromosome (YAC) clone that spanned the Pto region. Transcribed sequences within the Pto region were identified by isolating cDNA clones that hybridized to the YAC clone. Transformation of candidate cDNA clones into a Pst-susceptible, fenthion-insensitive tomato line succeeded in identifying a cDNA conferring Pst resistance and a separate cDNA conferring sensitivity to fenthion. The cDNA clones represent members of a tandemly repeated gene family and encode putative serine-threonine protein kinases. The role of these kinases in recognizing a signal from Pst or the fenthion molecule and in activating the plant response is currently being investigated.Abbreviations
Pst
Pseudomonas syringae pv. tomato
- RAPD
random amplified fragment polymorphism
- RFLP
restriction fragment length polymorphism
- YAC
yeast artifical chromosome 相似文献
4.
Summary Nucleus substitution of Brassica japonica (2n=20) with Raphanus sativus (2n=18) was carried out by means of repeated backcrossing of Brassicoraphavus (2n=37) to R. sativus as a pollen donor. In the course of nucleus substitution, chlorophyll deficiency appeared. Plants with more than 28 chromosomes, like their parents, had green leaves and those with 24 to 26 chromosomes had yellowish green ones. Almost all plants with 18 to 23 chromosomes showed yellow or whitish yellow. The R. sativus with B. japonica cytoplasm (2n=18) was obtained after four successive backerosses. The completely substituted R. sativus showed the same fertility as the true R. sativus used as a recurrent parent. It is assumed that the chlorophyll deficiency is caused by disharmony between the B. japonica cytoplasm and the R. sativus nucleus. The chlorophyll deficiency is discussed in comparison with male sterility or other characters which sometimes occur in alloplasmic Raphanus and Brassica species. 相似文献
5.
A. D. Munshi Bishwajit Panda Bikash Mandal I. S. Bisht E. S. Rao Ravinder Kumar 《Euphytica》2008,164(2):501-507
The genetics of resistance to Cucumber mosaic virus (CMV) in Cucumis sativus var. hardwickii R. Alef, the wild progenitor of cultivated cucumber was assessed by challenge inoculation and by natural infection of CMV.
Among the 31 genotypes of C. sativus var. hardwickii collected from 21 locations in India the lowest mean percent disease intensity (PDI) was recorded in IC-277048 (6.33%) while
the highest PDI was observed in IC-331631 (75.33%). All the four cultivated varieties (DC-1, DC-2, CHC-1 and CHC-2) showed
very high PDI and susceptible disease reaction. Based on mean PDI, 8 genotypes were categorized as resistant, 13 as moderately
resistant, 9 as moderately susceptible and one as susceptible. A chi-square test of frequency distribution based on mean PDI
in F2 progenies of six resistant × susceptible crosses revealed monogenic recessive Mendelian ratio 1(R):3(S) to be the best fit.
This monogenic recessive model was further confirmed by 1(R):1(S) ratio as the best fit for back cross with resistant parent
and no fit for either 3:1 or 1:1 in the back cross with the susceptible parent. The results revealed that CMV resistance in
C. sativus var. hardwickii was controlled by a single recessive gene. Considering the cross compatibility between C. sativus var. hardwickii and cultivated cucumber, the resistance trait can be easily transferred to cultivated species through simple backcross breeding. 相似文献
6.
用cDNA-AFLP技术构建基因组转录图谱 总被引：9，自引：0，他引：9
以分离群体mRNA反转录的cDNA为模板进行AFLP分析，在保留AFLP多态性丰富、稳定性高、无需了解序列信息等优点的同时，集中显示基因组表达序列的多态性差异，已逐渐发展成为基因差异表达显示、表达基因遗传连锁作图和基因克隆的常用方法。本文介绍了作图群体组配、总RNA提取与mRNA分离、cDNA合成、AFLP分析、转录图谱多态性分析作图等cDNA-AFLP分析的主要技术。 相似文献
7.
黏类小麦育性相关基因SSH文库的构建 总被引：6，自引：1，他引：5
为了深入研究黏类小麦雄性不育的分子遗传机制, 利用抑制差减杂交技术构建了黏类小麦育性相关基因的cDNA文库。文库质量检测表明, 差减杂交效率较高, 质量较好。在不育和可育cDNA文库中随机挑选120个阳性克隆进行测序, 获得100余条高质量的表达序列标签(EST)。对序列进行BLAST比对及功能注释, 比较了不育和可育cDNA文库的基因表达谱, 发现在可育cDNA文库中参与能量代谢的基因出现频率较高, 在不育cDNA文库中检测到了与花发育调控相关的MADS-box转录因子、细胞凋亡相关的泛素结合酶及阻遏淀粉合成的腺苷二磷酸葡萄糖磷酸化酶等相关基因。这些基因很可能与育性相关。 相似文献
8.
A novel and stable cytoplasmic male sterility CMS line of tuber mustard has been bred by subsequent backcrosses for 10 years.
Two specific markers atpA and orf220 were cloned and partially characterized in our previous study (Zhang et al. 2003). In this study, two new molecular markers, orf256 and orf305/orf324, have been isolated and identified. The orf256 gene size was found to be 825 bp in CMS line and a 1,357 bp in its maintainer line. Sequence analysis indicated that the
orf256 gene was an entire coding sequence and downstream of the cox1 gene. Interestingly, the 906 bp fragment, which contains part of the sequence of orf222, nad5 and orf139 genes, was found to be inserted from the 451st bp of 5′-flank of the 1,357 bp fragment. In the same way, the orf324 gene was isolated from CMS line and orf305 gene from its maintainer line. Both of them are entire coding sequences, upstream from nad3 and rps12 gene, and co-transcribed with the nad3 and rps12 genes. In addition, two molecular markers, orf256 and orf324/orf305, have been successfully converted into the SCAR markers. Subsequently, ORF256, ORF324, ORF305 protein and ORF256-M-431 fragment
are predicated to contain signal peptide sequences, and ORF220 was predicated to contain signal anchor sequence. RFLP analysis
results revealed that all of the molecular markers exhibited polymorphisms. Northern blot analysis indicated that the expression
level of these genes in CMS line is higher than that of the maintainer line. In the mass, all of these genes are expressed
lower in the leaf than that of floral organs between the CMS line and its maintainer line. The difference in expression pattern
of different mitochondrial specific marker genes suggests that the abundance of mitochondrial proteins is differentially regulated
in the organ/tissue development in tuber mustard. Results of this study also provide some novel and useful clues to explore
the biological function of these specific marker genes in the tuber mustard. 相似文献
9.
茉莉酸甲酯对中国红豆杉细胞基因表达的mRNA差异显示研究 总被引：5，自引：0，他引：5
茉莉酸甲酯是一种环戊烷衍生物型植物激素,在植物次生代谢物质生物合成中起着重要作用。我们以中国红豆杉悬浮培养细胞为材料,设两组平行实验,一组为阴性对照组,一组以茉莉酸甲酯进行诱导,所得细胞利用mRNA差异显示技术进行分析,研究了茉莉酸甲酯对红豆杉细胞mRNA表达的差异,最终共获得34个差异表达片段。经二次扩增和单引物阴性对照实验及序列测定,共获得9个阳性差异表达片段。与GenBank数据库比对,结果表明其中2个片段与已知基因有较高的同源性,与欧洲赤松（Pinus sylvestris）cDNA克隆的同源性为88％,与南方红豆杉（Taxus mairei）18SrRNA基因的同源性为93％,其余7个差异表达片段与GenBank中所有基因的同源性都非常低,可能代表了红豆杉细胞中的受茉莉酸甲酯诱导的未知基因序列。 相似文献
10.
Lu Xiao Bin Yi Yufeng Chen Zhen Huang Wei Chen Chaozhi Ma Jinxing Tu Tingdong Fu 《Euphytica》2008,164(2):377-384
7–7365AB is a recessive genic male sterile (RGMS) two-type line, which can be applied in a three-line system with the interim-maintainer,
7–7365C. Fertility of this system is controlled by two duplicate dominant epistatic genes (Bn;Ms3 and Bn;Ms4) and one recessive epistatic inhibitor gene (Bn;rf). Therefore an individual with the genotype of Bn;ms3ms3ms4ms4Rf_ exhibits male sterility, whereas, plant with Bn;ms3ms3ms4ms4rfrf shows fertility because homozygosity at the Bn;rf locus (Bn;rfrf) can inhibit the expression of two recessive male sterile genes in homozygous Bn;ms3ms3ms4ms4 plant. A cross of 7–7365A (Bn;ms3ms3ms4ms4RfRf) and 7–7365C (Bn;ms3ms3ms4ms4rfrf) can generate a complete male sterile population served as a mother line with restorer in alternative strips for the multiplication
of hybrid seeds. In the present study, molecular mapping of the Bn;Rf gene was performed in a BC1 population from the cross between 7–7365A and 7–7365C. Bulked segregant analysis (BSA) and amplified fragment length polymorphism
(AFLP) technique was used to identify molecular markers linked to the gene of interest. From a survey of 768 primer combinations,
seven AFLP markers were identified. The closest marker, XM5, was co-segregated with the Bn;Rf locus and successfully converted into a sequence characterized amplified region (SCAR) marker, designated as XSC5. Two flanking
markers, XM3 and XM2, were 0.6 cM and 2.6 cM away from the target gene, respectively. XM1 was subsequently mapped on linkage
group N7 using a doubled-haploid (DH) mapping population derived from the cross Tapidor × Ningyou7, available at IMSORB, UK.
To further confirm the location of the Bn;Rf gene, additional simple sequence repeat (SSR) markers in linkage group N7 from the reference maps were screened in the BC1 population. Two SSR markers, CB10594 and BRMS018, showed polymorphisms in our mapping population. The molecular markers found
in the present study will facilitate the selection of interim-maintainer. 相似文献
11.
Cytoplasmic male sterility (CMS) is a maternally inherited trait that fails to produce functional pollen grains. The CMS system
is widely employed to facilitate the utilization of heterosis in major crops. However, little is known about the CMS associated
genes in Upland cotton (Gossypium hirsutum). The objective of this study was to compare CMS cotton (CMS-D2) with the cytoplasm from G. harknessii and its isogenic maintainer line with the normal fertile Upland cotton cytoplasm to identify CMS-D2 specific gene(s) and
to develop CMS-specific sequence characterized amplified region (SCAR) markers. Based on Southern blot analysis using 10 mitochondrial
gene-specific probes (cob, cox2, atp6, atp9, nad3, cox3, atpA, cox1, nad6 and nad9), three probes (cox3, atpA, and nad6) revealed restriction fragment length polymorphisms (RFLP) between the CMS-D2 and its isogenic maintainer line. RT-PCR confirmed
that the three genes were differentially expressed between the two lines. These results indicated that there existed structural
and expression variations in the three genes when the mitochondrial D2 genome was transferred into Upland cotton. Genome walking
and rapid amplification of cDNA ends (RACE) were further performed to analyze the sequences of these genes and their flanking
regions. For cox3 and nad6, there was only one different nucleotide each in the gene regions between the two lines. Also some nucleotides upstream of
the ATG codon were different. For atpA, the sequences downstream the atpA were significantly different between the two cytoplasmic lines. Furthermore, two nucleotides at the -4 and -5 position from
ATG codon were also changed between the two cytoplasms (i.e., CG→AA), and this mutation also exists in RNA sequences. Interestingly,
nine nucleotides (ATGCAACTA) were also inserted at the location of 899 bp upstream of ATG codon in the CMS line. The results
suggest that the abnormal sequence and expression of atpA gene is associated with CMS expression in Upland cotton. According to the significant different sequences downstream the
atpA gene, a CMS-D2 specific SCAR marker was developed. The CMS-specific PCR bands were verified for 10 cultivars containing either
normal- or CMS-D2cytoplasm. This will allow quick and reliable identification of the cytoplasmic types of individual plants
at the seedling stage, and assessment of the purity of F1 seed lots. 相似文献
12.
Incompatibility in diploid and tetraploid crosses of Cucumis sativus and Cucumis metuliferus 总被引：1，自引：0，他引：1
The African horned cucumber (Cucumis metuliferus Naud.; 2x = 2n = 24) contains genes that can confer resistance to many important cucumber (C. sativus L.; 2x = 2n = 14) pests [e.g., root-knotnematode, Meloidogyne incognita (Kofoid & White) Chitwood]. Cucumber is highly susceptible to this root-knot nematode species, and a recent screening of
C. sativus accessions in the U.S. National Plant Germplasm collection did not identify sources of resistance. Thus,autotetraploids of
Cucumis sativus and C. metuliferus were created to recover fertile resistant interspecific progeny. Autotetraploids were obtained at the highest rate when seeds
were immersed in 0.5% colchicine for a period of 6 to 8 hrs. Treatment durations less than 6 hrs produced few tetraploids,
and durations of 10 hrs or more were lethal to seeds or developing seedlings. Crosses between C. sativus and C. metuliferus were made using diploid and tetraploid lines in all possible combinations, including reciprocals. Fruit development occurred
in crosses when diploid and tetraploid C. sativus were used as the female parent. However, seeds developed only in fruit of C. sativus (4n) ×C. metuliferus (2n) crossings. Seeds from these crosses, however,were flat and not viable. No fruit development occurred in crosses whereC. metuliferus was used as the female parent.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
13.
14.
不同优势抗虫棉杂交组合不同生育期基因表达差异初探 总被引：2，自引：0，他引：2
采用DDRT-PCR技术对3个产量优势差异较大的杂交棉组合及其亲本在4个生育期叶片cDNA进行扩增和差显,结果表明,产量高、中、低优势组合与其亲本基因差异表达比例从蕾期到花铃期总体上呈递减趋势;杂种特异表达在前3个生育期高优势组合高于中或低优势组合,特异表达可能对杂种优势的产生起一定作用;单亲表达一致是基因差异表达中最主要表达模式,但在各生育期高、中、低优势组合之间变化幅度较小,表明此模式可能与杂种优势发挥无明显关系。 相似文献
15.
J. R. Wall 《Euphytica》1967,16(2):199-208
Genetic control of sex expression in particular strains of Cucumis melo and C. sativus is under the control of a single locus. A dominant allele, A, is responsible for monoecious sex expression; the recessive allele, a, is responsible for andromonoecious sex expression. The evidence suggests that in C. melo fruit shape is determined by a single gene with incomplete dominance plus minor modifying genes, linked in coupling phase with the gene for determination of sex expression. In C. sativus the evidence does not completely exclude the possibility that the alleles for sex determination are pleiotropic and, therefore, are also the major determiners of fruit shape. However, evidence in favor of close linkage between genes determining sex and fruit shape is given and the author favors an interpretation of close linkage between two subunits of a complex gene. Evolutionary significance is seen in the presence of correlated inheritance of sex expression and fruit shape in three species of two genera of the Cucurbitaceae. 相似文献
16.
Summary
Atylosia scarabaeoides
Benth. and A. platycarpa
Benth., close relatives of the species Cajanus cajan (L.) Millsp., were screened for photoperiodic response. Four photoperiods ranging from 12 h 45 min to 19 h were studied in three environments. A. scarabaeoides flowered freely only in the first photoperiod. A. platycarpa not only flowered early (39 to 63 days after planting) in all four photoperiods, but also exhibited a relatively constant vegetative phase up to 16 h of illumination.
Cajanus cajan, in which most cultivars-if not all-exhibit a response to photoperiod, was successfully hybridized with the two Atylosia species. It is suggested that genes for earliness and insensitivity to day-length could be transferred from A. platycarpa to C. cajan. 相似文献
17.
Field evaluation of antisense RNA mediated inhibition of GBSS gene expression in potato 总被引：2，自引：0，他引：2
G. J. Kuipers J. T. M. Vreem H. Meyer E. Jacobsen W. J. Feenstra R. G. F. Visser 《Euphytica》1991,59(1):83-91
Summary Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Analysis of antisense RNA mediated inhibition of GBSS gene expression in large numbers of tubers from in vitro grown, greenhouse grown and field grown transgenic potato plants revealed stable and total inhibition of GBSS gene expression in one clone. In three other transgenic genotypes partial and unstable inhibition was found. In these genotypes both GBSS activity and amylose content were remarkably reduced compared with the non-transformed control genotype. No relationship was found between the level of inhibition of GBSS gene expression and yield and dry matter content. 相似文献
18.
We produced artificial interspecific chimeras by in vitro grafting, and obtained cytoplasmic male sterile (CMS) variants in the seed progenies derived from backcrossing the chimera with one of the mother plants, B. campestris cv. Komatsuna. The induced CMS has been stably inherited by crossing it with `Komatsuna', not with `Ruby Ball' cabbage. The nuclear component of CMS is complete `Komatsuna' type in morphology, chromosome number (2n = 20) and Southern blot using ribosomal 17S RNA gene as a probe. PCR analysis by using mitochondrial atpA primer showed the complete `Ruby Ball' type, suggesting nuclear-cytoplasmic exchange. However, Southern blot patterns were different among those of the CMS and both parents by using atpA. Recombination or some unknown change is supposed in the mitochondrial genome via the processes of synthesis and propagation of the chimeras. 相似文献
19.
Search for novel genes for resistance to powdery mildew (Sphaerotheca fuliginae) in cucumber (Cucumis sativus) 总被引：1，自引：0，他引：1
Summary The current powdery mildew (Sphaerotheca fuligninea) resistant cucumber varieties suffer from leaf chlorosis during autumn, winter and early spring cultivation in the Netherlands. Therefore screening was carried out for novel powdery mildew resistance genes. From 177 accessions, derived from different sources, 108 accessions proved to be partially resistant to S. fuliginea. Crosses were made with 53 resistant accessions to distinguish the presence of novel genes. It is likely that the accessions C. sativus 2145, C. sativus LV 41, PI 188807, Vladivostokij, White and Yellow 1 have one or more recessive powdery mildew resistance genes, different from powdery mildew resistance genes of the line NPI, which was used for variety breeding. Powdery mildew resistance tests with S. fuliginea give similar results in different regions of the world.Abbreviations pmr
powdery mildew resistance 相似文献