Bioinformation showed that BnRabGDI3 gene promoter included a 2 064bp-long segment at the upstream translation start site in Brassica napus genome. Promoter sequence contained several anther specific cis-elements. To verify the biology function, a recombinant vector was designated as pBnRabGDI3::GUS through the replacement of CaMV35S promoter in pBI121 by cloned BnRabGDI3 promoter fragment. In the vector, gus reporter gene was driven by BnRabGDI3 promoter. GUS histochemical assay in six independent transgenic plants showed that gus expressed only in anthers from the 11th to 13th development stages. The highest level of GUS activity was detected in bicellular and tricellar microspores, suggesting BnRabGDI3 played an important role in microspore development.
Aneuploidy Nj04-089 was an addition line derived from inter-specific cross of Brassica napus × B. carinata. To analyze the inheritance of the aneuploidy, chromosome of root-tip cell from two Nj04-089 consecutive-selfing generations were observed. Moreover, microscope and chromosome behaviors of pollen mother cells at meiosis were detected by PCR，Genomic in situ hybridization (GISH) and Southern blot. Results showed that two generations had chromosome numbers of 2n=37～51 in root-tip cells. The chromosome configurations of pollen mother cell at metaphaseⅠwere (0-4)Ⅰ+ (16-23)Ⅱ+ (0-2)Ⅲ and (0-8)Ⅰ+ (14-24)Ⅱ+ (0-4)Ⅲ+ (0-1)Ⅳ respectively. Abnormal behavior of chromosome pairing was detected including chromosome-bridge, chromosomes lagging, and chromosome unequal distribution. PCR，GISH, and Southern blot results showed that the aneuploidy had no genetic information on B genome. It could be concluded that the additional chromosomes were not from entire or large fragment of Brassica B genome.
According to the conserved region of PDAT gene available from GenBank, the cDNA sequence of PDAT gene from Jatropha curcas L. was cloned by degenerative PCR and RACE. The gene was named JcPDAT1 (GenBank accession HQ827796). The full length cDNA of JcPDAT1 is 2 869bp, encoding 670 amino acids. Sequence alignment and phylogenetic analysis revealed that JcPDAT1 shared high homology with PDAT from other plants, such as RcPDAT1 (Ricinus communis, 75%) and AtPDAT1 (Arabidopsis thaliana, 74%). The sequence harbored typical functional domains of PDAT. Realtime PCR showed that JcPDAT1 was expressed in different tissues including seeds, leaf, root tip, and was highly expressed in developing seeds, especially at the oil accumulation stage. TLC analysis and Nile red staining showed that JcPDAT1 could restore the TAG biosynthesis and accumulation in Saccharomyces cerevisiae mutant strain H1246???which indicated that the JcPDAT1 possessed PDAT enzyme activity.
To understand the function of xyloglucan endotransglycosylase/hydrolase (XTH) genes in Eruca sativa selfing affinity regulation in Eruca sativa, SC (self-compatibility) mutant and SI (self-incompatibility) wild type of E. sativa were used to clone DDRT (Differential Display Reverse Transcription) cDNA fragment. An XTH gene cDNA sequence (EsXTH1, 1074bp) was cloned from the cDNA of the whole flowers. The lengths of EsXTH1 5' end and 3' end untranslated regions were 36bp and 180bp. The length of open reading frame including 858bp coding a 285-amino-acids peptide. Deduced EsXTH1 protein sequence contains XTH protein active site DEIDFEFL. Compared to other species, EsXTH1 had higher homology to Arabidopsis thaliana AtXTH6. The homology to AtXTH7, SlXTH7 (tomato) and DcXTH4 (carnation) were 73%, 73% and 69% respectively. Molecular evolution analysis showed that EsXTH1, as well as AtXTH7, SlXTH7 and DcXTH4 were in the same group. EsXTH1 had XET and XEH active motifs. Its N-end had one signal peptide of 23 amino acids. EsXTH1 was putatively related to protein secretion and E. sativa fertility regulation.
Four different O/L peanut varieties were selected to analyze the relative quantitative expression of FAD2 gene in the different tissues by real-time fluorescence quantitative PCR technology. Results showed that FAD2 gene in different O/L genotypes presented constitutive expression. The expression of FAD2 gene were lower in the root, stem and leaves tissues. The expression in the flowers was significantly higher than that in pod and other tissues, and the difference among varieties was significant. The expression of HuaU606 and HuaU12 in flowers, young pods and mature pods tissues declined successively, and the expression of Huayu17 and Shiyouhong No. 4 showed high - low - high expression trends, the difference reached significant level.
More than fifty lines with segregated characteristics were obtained by crossing Glycine soja Sieb.et Zucc. and Glycine max (L.) Merr. to utilize and protect the Glycine soja, after selection over nearly three decades, 24 lines with mostly stable phenotypic traits were selected for this study. The leaf shape, flower color, growth habit, pod bearing habit, mature pod color, seed shape, seed coat color and hilum color were observed and described, with the male parent Glycine soja as control. These traits showed abundant variation among all lines. Branch number, plant height, main stem diameter, yield per plant and 100-seed weight were then measured, and the coefficient of variation was as follows: 100-seed weight > yield per plant > plant height > branch number > main stem diameter. All lines were divided into 4 groups using McQuitty’s Cluster analysis combined with the utilization characteristics. Among them, 5 lines were to be used as seeds, 6 (7 in Chinese above?) lines as grazing plant and 2 lines as both seeds and grazing plants.
Dry seeds of peanut cultivars Huayu 22 were irradiated by 14 MeV fast neutrons with dosages of 0, 9.7, 14.0 and 18.0Gy respectively. Embryonic leaflets were separated from the irradiated seeds and incubated on MSB5 induction media containing 10mg/L 2,4-D. After 4 weeks culture, the subcultures were transferred to MSB5 media supplemented with 4mg/L BAP. The results showed that the frequencies of somatic embryo formation and plantlet regeneration varied with different dosages of fast neutron irradiation. The somatic embryo induction rates and plant regeneration rates declined with the increase of irradiation dosages. The optimum irradiation dose of fast neutron for Huayu22 was between 9.7Gy and 14.0Gy. The regenerated plantlets were sliced, grafted onto peanut seedlings of Huayu 23 and transplanted into field. The plants grew to maturity and produced viable seeds.
）
Genetic diversity of 73 mustard (Brassica juncea L.) accessions from Tibet Autonomous Region of China and 35 from southwest of China，northwest of China and India were analyzed using 8 SRAP (sequence－related amplified polymorphism) primers combinations on 20 phenotypic traits. SRAP results showed that a total of 139 bands were produced with polymorphism rate of 24.5%. Clustering result of SRAP showed that 108 accessions were divided into 4 groups, and the accessions in each group were mainly from the same region. Genetic diversity index of China Tibet accessions was higher than others. Variation degree of 8 important phenotypic traits of Tibet accessions showed the highest variation coefficiency was 41.29% on silique number per plant. Clustering analysis of phenotypic traits showed that 108 materials were clearly divided into different groups based on their original regions except the accessions from Northwest. In conclusion, Tibet mustard had abundant genetic diversity. The order of genetic diversity indexes from top to bottom was as follows: China Tibet, Southwestern China, Northwestern China and India. Mustard genetic diversity was mainly associated with geological and biological conditions.
Lack of available phosphorus in Southern acid red soils was one of the major limitations to soybean production. Breeding phosphorus tolerant varieties was one of the most efficient ways to improve production . Using high and low phosphorus soil pot experiment, the phosphorus efficiency of 5 Brazilian soybean and their 11 derived varieties were evaluated using 10 traits which were shoot dry weight, root dry weight, shoot phosphorus content, root phosphorus content, plant phosphorus uptake, root length, root surface area, root volume, phosphorus use efficiency and phosphorus absorption efficiency. Furthermore, phosphorus efficiency of 19 soybean varieties were analyzed by principal component and associated functions. The results showed that except Brazil 13 and Brazil 9, phosphorus efficiency traits of the other Brazilian soybean resources were generally better than Guizao 1 which was a variety in South China. Huaxia5 and Guixiadou2 varieties were higher than their parents respectively, which was a positive transgressive inheritance. Huaxia1 and Huaxia 3 varieties were between their parents. The remaining varieties were lower than their parents respectively, which was a negative transgressive inheritance. It indicated that the majority of Brazilian soybean resources were superior to South China varieties in phosphorus efficiency. Therefore, Brazilian soybean resources had great potential to improve South China varieties of phosphorus efficiency.
In order to understand the changes of leaf nitrogen content and its relationship with the net photosynthetic rate (Pn) during genetic improvement of soybean cultivars, leaf nitrogen content and photosynthetic characteristics of 21 Soybean cultivars released from 1923 to 2005 in Jilin Province were studied. The results showed that the leaf nitrogen content had highly significant or significant positive correlation with released years in R2 and R6 stages, but it had no significant negative correlation in R4 stages. The growth rate of nitrogen content per leaf area was significantly higher than the net photosynthetic rate in R2 and R6 stages, resulting in highly significant or significant negative correlation between photosynthetic nitrogen efficiency and released years. But the nitrogen content per leaf area of modern cultivars was lower than the old cultivars in R4 stages, the net photosynthetic rate was still higher than the old cultivars, resulting in a significant positive correlation between photosynthetic nitrogen efficiency and released years. The chlorophyll content and the net photosynthetic rate would be high if the nitrogen content in leaf was high, but the photosynthetic nitrogen efficiency would be low. In R4 stages, the photosynthetic nitrogen efficiency could be a high-yielding cultivars bred indicator.
For rapeseed mechanization production, effects of rape density and nitrogen (N) application rate on growth and N use efficiency were investigated under field condition. Two planting density (15?104 and 45?104 plant per hectare) and four N rates (0, 90, 180 and 270kg per hectare) were applied to study the interaction between N and density on agronomic characteristics, yield, N use efficiency and N fertilizer requirements for Zhongshuang 11. Results showed that plant height, branch number, effective siliques per plant and seeds per pod significantly decreased at higher density, but branch height and 1 000-seeds weight increased significantly. Under higher density, the plant-type was uniform and ideal for mechanized harvesting. Yield of high density rape was higher. Both yields on high and low density increased at 0 to 180kg/hm2 N rates. But when N rates were up to 270kg/hm2, both yields decreased. N use efficiency decreased at higher N rates and lower density. It indicated that higher density could promote seed N accumulation, decrease N fertilizer requirement and improve the N use efficiency. For the same targets yield (compared to the top economic yield at low density, the average yield of Hubei Province and that of China), the N fertilizer requirement significantly decreased by 22.9% to 30.6% after increasing planting density.
Key words: Rapeseed (Brassica napus L.); Planting density; Nitrogen fertilizer; Yield; Nitrogen use efficiency
Plot experiments were carried out to study the effects of nitrogen (N) application rate on fresh yield, nutrient contents and accumulations, seed yield and N use efficiency of Orychophragmus violaceus. Results showed that compared to no N application, the fresh yield, nutrient accumulations and seed yield were increased significantly after N application. With 135 kg/hm2 N application rate, fresh yield, N, P, K accumulations and seed yield were optimal, reaching 21350 kg/hm2, 67.17 kg/hm2, 11.62 kg/hm2, 87.05 kg/hm2 and 993 kg/hm2 respectively. With increased N application rate，N agronomic efficiency (AUE), partial factor productivity ( PFP) and apparent recovery efficiency ( ARE) of Orychophragmus violaceus decreased significantly. Excessive N had negative effects on growth of Orychophragmus violaceus.
Household survey was carried out by soil testing formulated fertilization project from 2006 to 2009 in Qin-Ba mountain area of southern Shaanxi. The object was to evaluate current situation of fertilizer application and problems of nutrients input by households on rapeseed in this region. Data was collected from 2 576 households in11 counties. Results showed that in this area, the average yield of rapeseed was 2 355kg/hm2 over 60.68% households. The rates of total N, P2O5 and K2O were 179, 80 and 54kg/hm2 respectively. Among them, the chemical fertilizer rates were 145, 62 and 34kg/hm2 respectively. Yield had significant correlation with chemical fertilizers. Diminishing return was found in each fertilizer input. According to yield level classification, households with rational chemical fertilizer (of N, P2O5 and K2O) rates were 38.55%, 27.60% and 25.89% respectively, while households with excessive application were 15.22%, 26.24% and 10.33% respectively, and households with insufficient application were 46.23%, 46.16% and 63.78% respectively. The survey showed that, if the chemical fertilizer was supplied at a rational level, the yield would increase 56.1 thousand tons. Using boron fertilizer would increase 101kg/hm2 in yield. Moreover, households using organic and boron fertilizer were only 45.26% and 41.73% respectively, which could further increased. The problems of fertilizer input in this region were irrational nitrogen and phosphate inputs, as well as insufficient potassium, boron fertilizer and organic manure input. It was recommended to balance the nitrogen and phosphate application, increase potassium and boron fertilizers, increase organic manure inputs, and increase top dressing fertilizers, especially potassium.
The pathogen causing browning of vascular system and pith of soybean in China was identified using morphological and molecular characteristics. Colonies of the isolates on PDA medium developed in the form of a flat, dense, roughly circular, white to putty-coloured or grey mat, with radial folds and a variable, usually finely lobate or notched margin. Colonies on GBA medium were white to grey, circular, with smooth edge whose mycelia adhered to medium and the aerial mycelia were sparse. On GBA medium, the conidia were produced, which are hyaline, aseptate, uniseptate or multiseptate, ovoid to elliptical, (3.0 ~ 5.1) ×(2.1 ~3.3)?m. Typical symptoms of brown stem rot occurred in 100% of seedlings when inoculated with the isolates on soybean cultivar Hefeng 25. To verify the pathogen, rDNA-ITS fragments of the isolates amplified using primers ITS1 and ITS4 were sequenced, and 99% of sequence similarity reached between the isolate and a Phialophora gregata isolate (GenBank Accession No. U66728.1) in Genbank. The specific 483-bp DNA products were amplified from the isolates using primers BSR1/BSR2 specific to P. gregata. The above results indicated the isolates were P. gregata.
In this study, 408 soybean cultivars (lines) were identified for resistance to the strong virulent strains of Fusarium oxysporum and F. solani from northeast China. Results showed that the resistant resources of soybean against F. oxysporum and F. solani were abundant in northeast China. Among 357 soybean cultivars (lines) from Heilongjiang Province, resistant cultivars against F. solani and F. oxysporum accounted for 39.03% and 62.37% respectively. Among 27 soybean cultivars from Jilin Province, resistant cultivars against F. solani and F. oxysporum accounted for 23.46% and 46.91% respectively. Among 24 cultivars from Liaoning Province, resistant cultivars against F. solani and F. oxysporum accounted for 25.00% and 61.11% respectively. Jiaojin05-SH, Heinong65, D09-005, D09-072 and D09-077 from Heilongjiang Province showed resistance against strong virulent strains of F. solani and F. oxysporum from northeast China. There were differences in the virulence of F. solani and F. oxysporum strains from different provinces to the same soybean cultivar (line), so the local Fusarium strain should be selected for resistance identification. Ten major soybean cultivars from northeast China were identified for resistance against eight Fusarium species with strong and weak virulence. The results showed that resistant interactions accounted for 74.38% among the 160 interactions, indicating that the resistance against Fusarium root rot existed abundant in soybean cultivars. Suinong28 and Dandou13 were immune to weak virulent F. avenaceum, Heinong44 and dongnong2037 immunized to weak virulence F. oxysporum; and at the same time there was no cultivar resistant to all eight Fusarium species. According to the proportion of resistance interactions and the average disease index, the resistant frequency of ten cultivars descending order were Jiunong28, Jiyu35, Suinong28, Tiedou44, Heinong44, Dandou13, Kenfeng16, Dongnong2037, Liaodou17, Hefeng55. Cultivars shown resistance to both Fusarium strong and weak virulence strains accounted for 71.25%, therefore, virulence of Fusarium strain was not taken into account for identifying resistance of soybean cultivar to Fusarium species.
Maize (Zea mays L.) and soybean [Glycine max (L.) Merr.] are frequently grown in rotation throughout Northeast China. Soybeans injury often occurs when soybeans are grown in fields treated with atrazine [2-chloro-4, 6-bis-(ethy1amino)-s-triazine] the previous year for control of weeds in maize. Pot culture experiments were conducted to examine 45 soybean varieties with various seed size (seed weight range 81﹣302 mg/seed) for the response of soybean shoot and root injury from atrazine in the greenhouse. The results showed that dry shoot weight production of soybean grown in soil treated-atrazine generally increased significantly (P＜0.05) as seed size increased, while root dry weight production of soybean did not increased nonsignificantly (P＞0.05) as with larger seed size increased. The results indicated that large-seeded varieties of soybeans are more tolerant ofted atrazine better than small-seeded varieties. We suggest soybean injury from atrazine may could be reduced by planting large-seeded varieties or planting the largest seed from a given variety.
To explore the MOE (margin of exposure) application in risk assessment of peanut aflatoxins in consumers, data of aflatoxins contamination in post-harvest peanut of China and the data of Chinese peanut consumption were collected. Based on aflatoxins toxicological data, MOE values of different regions from population of different age-groups were evaluated. Results indicated no significant difference between urban and rural regions, but children were at higher risk than other populations. Compared to low-dose extrapolation, MOE was recommended due to its effective performance in posterior analysis.
Effects of salinity on the growth of Phaeodactylum tricornutum and content of organics in P. tricornutum cells were investigated. The culture condition of high yield of algal oil was also optimized. Results showed clear effects of salinities on biomass and accumulation of organics. The algal growth rate under 70‰ (?) salinity was 22.6% lower than that of normal salinity (35‰). The highest biomass (dry weight 1.92g/L) and total lipid (29.98%) were produced under normal sea water cultures. The highest volumetric productivity of total lipid under 35‰ salinity was significantly different, with the algal oil productivity of 47.67mg per litre each day. Polysaccharides content of dry weight reached up to 5.5% under 17‰ salinity, and protein reached up to 17.82% under 70‰ salinity.
The expression pattern of soybean class III acidic endochitinase gene in different soybean tissues was detected by real-time quantitative PCR (RTQ-PCR). Results showed this enzyme activity was lower in roots, stems and leaves, but higher in flowers and seeds. 5’-flanking upstream 2000bp sequence of soybean class III acidic endochitinase gene was isolated from soybean genomic DNA by PCR, and named CP. Sequence analysis revealed that this fragment contained a series of motifs related to seed-specific promoters and some pollen-expressed elements, such as SEF4 motif, E-box, G-box, (CA)n, AACA, ACGT, CCAA, 52-box, ntp303-box, GTGA, TACPyATbox. It was inferred that CP promoter could drive downstream gene over-expression in soybean flowers and seeds.
DNA methylase in plants plays an important role on maintaining genome DNA methylation, regulating gene expression and prompt response to varied adverse circumstances. Many bioinformatics websites and softwares were employed in this paper. Four kinds of DNA methylases and corresponding gene orders were determined preliminarily in soybean. Their evolutionary relationships and gene copy numbers were analyzed in this study. The locations of all genes on the chromosomes and the physio-chemical property and subcellular localization of DNA methylases were identified. The domains of these proteins and tissue-specificity expression were identified. These results provided a guidline in further understanding the characteristics of soybean DNA methylases and their genetics.
