Yi chuan xue bao = Acta genetica Sinica最新文献

The cytoplasmic male-sterility (CMS) was transferred into leaf mustard by inter-varietal hybridization and subsequent backcrosses cms donor as using tuber mustard. Using specially designed primers, one specific band at 600-700 bp was distinctively amplified by PCR from cms plants of different backcrossed generations. It was genetically stable and cytoplasmically inheritable in the various backcrossed progenies. The sequencing result shows it is a 663 bp fragment with its own initiation and termination codons, presumably encoding for 220 amino acids (named orf 220). This deduced polypeptide had two trans-membrane hydrophilic domains and its N-terminus shared high similarity in amino acids constituents with COX III protein from Oenothera berteriana, ATP8 protein from radish and ORFB protein from sunflower. In addition, RT-PCR analysis showed, orf 220 gene was constitutively expressed in leaves, roots and floral buds. However, the direct evidence of its involvement in expression of cms trait needs further confirmation.

In a recombinant inbred line (RIL) population of indica rice, two subpopulations composed of susceptible lines were selected for mapping of partial resistance to leaf blast with two isolates of the pathogen. A third subpopulation composed of susceptible lines with similar heading time was used for mapping of partial resistance to neck blast with a third isolate. The traits measured for partial resistance included diseased leaf area (DLA), lesion size (LS) and lesion number (LN) for leaf blast and lesion length (LL) and conidium amount (CA) for neck blast. A linkage map consisting of 168 DNA markers was constructed by using the whole RIL population. Quantitative trait loci (QTLs) conditioning these traits were determined at one-locus and two-locus levels. Eleven main-effect QTLs and 28 digenic interactions were detected by QTLMapper 1.01 b. Only three QTLs showing main effects were also involved in digenic interactions for the same trait. General contributions of epistatic QTLs of each trait ranged from 16.0% to 51.7%, while those of main-effect QTLs of each trait ranged from 4.7% to 38.8%. The general contributions of main-effect QTLs of most traits were smaller than those of epistatic QTLs, confirming the importance of epistasis as the genetic basis for complex traits. The general contributions of the main and epistatic effects of all QTLs detected for the two traits LL and CA of the partial resistance to neck blast reached 70.6% and 82.6% respectively, which obviously represented a major part of the genetic basis controlling partial resistance to neck blast. The results indicated the necessity for partial resistance mapping to use susceptible subpopulations where the interference of major resistance genes is avoided.

On the basis of the molecular linkage map, mapmaker software QTLMapper 2.0 was used to analyze the QTLs effect of the whole cocoon weight,cocoon shell weight, ratio of cocoon shell and pupa weight of domestic silkworm. For these four cocoon quantitative traits, 7, 6, 2 and 8 effective QTLs were detected and mapped to 7, 5, 2 and 7 linkage groups, respectively. Complicated epistatic effects were found involved in the genetic variation of the whole cocoon weight and cocoon shell weight. For the whole cocoon weight, there were three pairs of QTLs with significant additive by additive interactions, in which, one pair had significant additive by dominance and dominance by dominance interactions. Whereas significant dominance were detected for three QTLs and significant additive effects one QTL had. For the cocoon shell weight, significant genetic effects, including epistatic effects were found for one pair of QTLs, significant dominance by dominance interaction for another pair of QTLs; one QTL had significant dominance and another QTL had additive by additive interaction. The ratio of cocoon shell and the pupa weight were controlled mainly by additive or dominance effects. No interaction between QTL was found for the ratio of cocoon. Most QTLs, associated with the pupa weight, had negative dominance effects. Only significant additive by additive interaction was found between one pair of QTLs. The 2nd, 3rd, 4th, 11th, 13th, 24th, 34th, 37th, and 40th linkage groups are the common chromosomal regions harboring QTLs of two or more cocoon quantitative traits. There are identical QTL or chromosomal region for the whole cocoon weight and cocoon shell weight, indicating they can be simultaneously improved by utilizing epistatic effects in breeding.

Nucleophosmin (NPM) is an abundant nucleolar phosphoprotein. NPM gene involved chromosomal translocations were found in the patients with anaplastic large cell lymphomas (ALCL), myelodysplastic syndrome (MDS), acute myeloid leukemia (AML) and acute promyelocytic leukemia (APL). To generate NPM gene knockout mice and study its biological function in vivo, we screened the lambda phage genomic library derived from 129S1 mice with mouse NPM cDNA probe. A positive phage clone which contained the full-length NPM genomic DNA was obtained and the insert of 15.3 kb genomic DNA in this clone was sequenced with shotgun method. BLAST analysis showed that the sequence of insert are 99.8% identity to that of NPM gene of C57BL/6 mouse strain. Based on the sequence, bioinformatics analysis on genomic structure of NPM and the transcription factor binding sites in the NPM 5' flanking region were performed.

A system of virus-induced post-transcriptional gene silencing for studying rbcS gene function was established and optimized using tobacco rattle virus vector and Nicotiana benthamiana as experimental materiaes. The following analyses were conducted: phenotypic characterization of rbcS gene silenced plants, transcription levels of rbcS gene by RT-PCR; protein levels of rbcS by the antibodies of rbcS and rbcL and photosynthetic pigments wntents in rbcS silenced plants by HPLC method. The results showed that the seedlings at 21-24-day-old and Agrobacterium concentration at OD600 = 1-1.5 gave the best results for gene silencing. The expression level of rbcL was very likely regulated by rbcS, and rbcS gene did not relate to the collection of photosynthetic energy. Probability analysis showed that the tobacco rattle virus vector system is a useful and effective technique to study rbcS gene function via post-transcriptional gene silencing.

One 134 bp fragment was amplified in anthers of male sterile and fertile wheat using one pair of degenerated primer designed based on the conserved domain of MS2 gene in Arabidopsis thaliana and Oryza sativa, and one 1604 bp male sterility gene homology sequence was extended by in silico cloning based on the 134 bp fragment. The amino acids encoded by the male sterility gene homology sequence include a 200 amino acid conserved domain of male sterility, and this sequence expressed only in wheat male fertile anthers; no expression in male sterile anthers, roots and leaves. This research demonstrated that the cloned male sterile homology sequence is specific to wheat anther development.

SMAD3 is one of the receptor-activated SMADs which are important in TGF-beta signal transduction. Smad3-null mice show accelerated cutaneous wound healing compared with wild-type mice. In this work, we investigated the functions and the mechanism of Smad3-mediated TGF-beta signal on matrix metaloproteinase-2 (MMP-2) in mouse fibroblast. We found that MMP-2 at wound bed was expressed earlier in Smad3-null mice than that in wild type and heterozygotes. In the sera of wounding mice, the activity of MMP-2 was also remarkably higher in Smad3-null mice than that in the other two. The embryonic fibroblasts were separated from Smad3 knockout mice to test the function of Smad3 in modulating the expression of MMP-2. The results showed that the expression and activity of MMP-2 in Smad3-null fibroblasts were higher than those in wild type cells. TGF-beta1 could increase the MMP-2 activities in both Smad3 mutant and wild type fibroblasts. The expression and activity of MMP-2 were inhibited by over expression of SMAD3 in Smad3-null fibroblasts, while the expression and activity of MMP-2 were increased by over expression of anti-sense Smad3 in wild type cells. All these results showed that SMAD3 inhibited the expression of MMP-2 in mouse embryonic fibroblasts.

The embryo-derived calli from four types of tall fescues (Festuca arundinacea Schreb) were transformed with two Agrobactrium tumefaciens strains AGL1 and GV3101. AGL1 harbors an intron-AtNHX1 expression vector pROK2U containing ubiqutin promoter and npt II marker gene. GV3101 harbors an intron-AtNHX1 expression vector pROK2 containing 35S promoter and npt II gene. After infection and co-culture with AGL1 or GV3101, the embyogenic calli were selected with 50-150 mg/L paromomycine and 1126 resistant plants regenerated from the resistant calli. All plants were selected further with 10-20 mg/L Kanamycin and 525 of them remained green. Genome DNA of the resistant plants was checked with specific primers and probe from AtNHX1 gene. The results of PCR assay and Southern blot analysis indicted that exogenous target gene (AtNHX1 gene) had been transferred into different cultivars of Festuca arundinacea. Different transformation frequencies among the four cultivars were obtained.

We analyzed the electrophoritic patterns of 16 kinds of isozyme and three kinds of storage protein in Agropyron elongatum 2x and Ag. elongatum 4x by isoelectric focusing (IEF) and SDS-PAGE, respectively. The results showed three types of electrophoritic patterns. In the first type, the zymogram phenotypes are identical between Ag. elongatum 2x and Ag. elongatum 4x. Two kinds of isozyme belong to this type, which accounts for 10.5% of all the biochemical markers analyzed. In the second type, Ag. elongatum 4x showed a phenotype which is made up of combinations of the all of Ag. elongatum 2x bands and the specific bands. Ten kinds of isozyme and three kinds of storage protein fall into this class, which is the most part and accounts for 63.2% of all the markers analyzed. In the third type, Ag. elongatum 2x and Ag. elongatum 4x showed some identical bands and their own specific bands. Five kinds of isozyme were classified to this group,which accounts for 26.3% of all the markers analyzed. The results above suggested that Ag. elongatum 4x is a allotetraploid composed of one genome originated from Ag. elongatum 2x and another unknown genome which is apparently distinct from the St, J and N genomes of relative species. Otherwise, SSR primers were used to analyze the relation of Ag. elongatum 2x and Ag. elongatum 4x. The amplification results of most primers showed that Ag. elongatum 2x and Ag. elongatum 4x have some identical bands and Ag. elongatum 4x have some specific bands. This result validated the conclusion from biochemical marker analyses that Ag. elongatum 4x is a allotetraploid.

Agrobacterium-mediated transformation is probably the most widely used method to introduce genes into plants. Great progress has been made in recent years in studies on the mechanism of Agrobacterium-mediated transformation. Agrobacterium genetically transforms plants by transferring a portion of the resident Ti-plasmid, the T-DNA, to the plant. VirD2 and VirE2 accompany the T-DNA into the plant cell. Both proteins may aid in T-DNA transfer, nuclear targeting and integration into the plant genome. In recent years, some Arabidopsis rat (resistant to transformation) mutants are deficient in T-DNA integration according to some studies. These results showed that plant genes participate in the T-DNA transport and integration process. This paper discusses our current knowledge about the functions of virulence protein, namely VirD2 and VirE2, and plant genes in several aspect of Agrobacterium transformation. The paper discusses two different classes of integration patterns in detail: one is T-DNA right border being linked to genomic sequences by the VirD2 protein, the other is T-DNA integration via SDSA (synthesis-dependent strand-annealing). According to the latest studies we elaborated the T-DNA integration model based on genomic DSB (double-strand breaks) and proposed a new opinion about the mechanism of T-DNA integration.