To investigated fatty acid changes of a down-regulated fad2 gene transgenic rapeseed line W-4, its T5 to T7 generations and wild type Westar (WT) were used to study seeds fatty acid composition. Results showed that the increase of oleic acid and eicosenoic acid contents were highly significant. Oleic acid was 84.62%±1.41% in W-4 seed, 25.91% higher than that of wild type and eicosenoic acid was 18.46% higher too. While linoleic acid and linolenic acid were 3.22%±0.56% and 3.45%±0.51% respectively, which were 80.89% and 47% lower than that of WT respectively, and the decrease were highly significant. Moreover, palmitic acid (-18.10%) and stearic acid (-8.33%) were lower than that of WT, and the difference were highly significant. The results indicated that down-regulation of fad2 gene significantly changed the fatty acid composition. Thus the polyunsaturated fatty acid contents were significantly reduced, and monounsaturated fatty acid contents were increased.
A highly resistant germplasm ICGV86699 and a highly susceptible cultivar Zhonghua 5 were used as parents to build a recombined inbred lines population (XA-RILs). The genetic characteristics of resistance to late leaf spot (LLS) in peanut were studied based on the segregation analysis of genetic system of quantitative traits. The results showed that LLS resistance among XA-RILs was significantly different and the resistance was controlled by two additive-epistatic major genes and additive-epistatic polygenes. The heritabilities of main genes were 60.10%-86.61% while the heritabilities of polygenes were 6.65%-32.77%. The correlation analysis verified that resistance to LLS was linked with some poor agronomic traits such as late maturity, low yield and small seed. From XA-RILs, an elite line, XA006, was selected as a breeding line possessing high yield and resistance to LLS with other desirable agronomical traits.
Increasing the sulfur-containing amino acid content has been one of the most important goals to improve soybean quality. The objectives of this paper were to discover the soybean sulfur-containing amino acid Meta-QTLs and candidate genes related to sulfur-containing amino acid components in order to establish a basis of developing functional markers for the sulfur-containing amino acids. 33 QTLs related to sulfur-containing amino acids in soybean were integrated by using the reference map of Consensus Map 4.0 high density soybean genetic map. By Meta-analysis method, 8 Meta-QTLs for sulfur-containing amino acids were estimated. 7 long sequences used for screening candidate genes were identified in Meta –QTLs, by collinear interval of QTL sequence. 16 sulfur-containing amino acid related genes were acquired by using function annotation of genes, including 6 genes of amino acid synthetic enzyme, 1 gene of 11s glycinin A5A4B3 and 9 regulated genes. Regulated genes respectively encoded F-Box protein, PPR protein and transcription factors. There were EST profiles and SNPs of 13 genes in NCBI database. Analysis of candidate genes related to sulfur-containing amino acids in soybean could provide useful information for molecular marker assisted breeding.
The coat protein (CP) of Soybean mosaic virus (SMV) is the SMV genome structural proteins, it plays an important role in the process of SMV infection soybean. In this study the coding sequence of the CP in SMV was analyzed with Codon W and CUSP（Create a codon usage table）programs while compared with the genome of SMV. The results showed that CP of SMV was biased toward the synonymous codons with A and T. Compared with the codon usage of SMV genome, Codon Usage bias of the CP is the same as eight amino acid codon in the genome of SMV. The correlation analysis between ENC and GC3 indicated that the codon usage of CP gene was affected by many factors including the nucleotide composition. The cluster analysis results based on the gene CDS sequences showed codon usage bias of the CP gene were similar to the bias on the relative synonymous codon usage（RSCU）. The analyzing of the CP codon usage characteristics provided a foundation for further study its functionality.
12 MG standard varieties of MG0～MGⅢ from north USA sown in Sanjiang plateau of Heilongjiang were used as reference to identify the MG categories for local popular varieties in Heilongjiang in 2012 and 2013. The results showed that the maturity group’s growth period of the north USA standard varieties presented constant increase trend, and the borders were obvious that can be used for maturity group classification. There were significant differences in different growth period between group varieties and standard varieties. Results from 2 years showed that Huajiang 2, Heihe 38, Heihe 43 and Heihe 45 belonged to MG0 group; Hefeng 51, Hefeng 50, Hefeng 55, Suinong 26, Suinong 28 and Heinong 44 belonged to MGⅠgroup; the varieties of Kenfeng 16 and Heinong 51 belong to MGⅡ group.
SnRK2 genes encoding Ser/Thr kinase exist exclusively in plants. To understand SnRK2 function responding to abiotic stresses in castor bean, 6 SnRK2 genes were identified and characterized based on complete genome data of castor bean. Results showed that SnRK2s were hydrophilic proteins and their gene sequences were highly conserved in plants. Phylogenetic analysis revealed that SnRK2s were significantly differentiated between monocotyledons and dicotyledons, implying their different evolution history. High-throughput RNA-seq data were used to define the expressional profiles of SnRK2s among tissues and different development stages of seeds. Results showed that most of SnRK2s expressed in all tested tissues and tissue specific expression was not detected. But expressions of Group 2 genes were significantly up-regulated by stress. Among them, SnRK2 gene 29908.m006067 responded specifically to exogenous ABA(100μmol/L), NaCl (250mmol/L) and cold (4℃) treatments in seedlings. And gene 29772.m000313 responded specifically to ABA and cold.
To better understand the variations related to winterness and springiness of Brassica rapa, 4 BrFLCs genes from 5 turnip rape cultivars (different in temperature sensitivity and cold hardiness) were investigated by nested and semi nested PCR. Primers were designed between the sequence of exon 4 to exon 7 in the BrFLCs. Results showed that BrFLC1, BrFLC2 and BrFLC3 were amplified except BrFLC5. Sequence alignment analysis found some variations in the regions from exon 4 to 7 of the 3 BrFLCs. The main allelic variations were SNPs and Indels. Combining with their temperature sensitivity, variations in BrFLC1 had little correlation on to winterness, and those from BrFLC2 and BrFLC3 might have close higher correlations. 1 Indel and 5 SNPs in BrFLC2, and 1 Indel and 16 SNPs in BrFLC3 might be associated with winterness.
Initial blooming is the key stage of rapeseed growth. To understand the effect of drought on rapeseed at this stage, 10 days of drought (beginning from the day when field moisture was 45-50%) was conducted in pots under rain-proof shelter during initial flowering period. Normal irrigation was controlled (with 80-85% field moisture). 4 cultivars were used, including 2 drought-resistant Brassica napus L. cv Xunyou 8 and Yanguang 2009, and 2 drought-sensitive cv Zheyou 50 and Huayouza 13. After 10d-drought, resistant cultivars’ leaf SPAD value, soluble protein and malondialdelyde (MDA) content changed little, soluble sugar content declined slightly, catalase (CAT) activity increased slightly, while free proline significantly increased by 2.9-4.1 folds. On the contrary, sensitive cultivars’ physiology indicators changed significantly, including decreased SPAD value, soluble protein and soluble sugar contents, and increased free proline (but less than resistant cultivars). The drought stress had significant impacts on agronomic traits and yield. For sensitive cultivars, plant height, primary branches, inflorescence length, pods number on main inflorescence, pod length, pods per plant, seeds per pod and yield decreased significantly. But for resistant cultivars, the decrease rate was much lower.
To understand the early maturing mechanism of rapeseed (Brassica napus L.) variety, 3 materials [early matured Zhongyou 16 (ZY116), mid-early matured L01 and late matured Zheshuang 8 (ZS8)] were selected and their key enzymes of carbon and nitrogen metabolism were investigated. Results showed that ZY116 had lower leaf glutamine synthetase (GS) activity during initial flowering stage and lower grain sucrose synthase (SS) activity during late podding period than those of L01. But it had higher leaf sucrose phosphate synthase (SPS) activity during initial flowering stage and grain glutamate synthase (GOGAT) activity during initial podding stage than those of L01. The max C/N ratio during budding stage was different among the varieties. The peak of C/N ratio before flowering stage was lower than that of L01.
Root morphology characteristics, physiological activity and biomass accumulation were studied to investigate the root response to shading under relay-intercropping system in seedling stage. 3 varieties with different root characteristics, NTS1007, GX-3 and ND12 were used in the pod experiment to investigate root activity, root surface area （RSA）, root active absorption area （RAAA）, bleeding sap and biomass accumulation. Compared with the monoculture, the results showed that the length of main root and lateral root decreased significantly in the relay strip intercropping system, and the increase in lateral root （81.8%） was more obvious than that of main root （42.8%）. The soluble sugar content, root activity, bleeding sap, root surface area, and root active absorption area all decreased significantly. The decreasing amplitude of root surface and active absorption area were linearly correlated with the first lateral root length. Biomass decreased and the decrease in the root biomass （55.2%） was more apparent than shoot （37.9%）. Among the 3 varieties, ND12 had the least decrease in main root length, first lateral root length and root volume （33.9%, 74.4%, 65.3%）, but GX-3 had the most significant decrease in RSA and biomass （67.1%, 48.3%） in the relay strip intercropping system, and NTS had the greatest decreasing aptitude in R/S, root activity and bleeding sap/shoot dry matter （35.4%,39.2%,38.9%）. In a relay strip intercropping system, the elongation growth of the soybean root can reflect the status of the root growth more accurately in comparison to monoculture, which can be used as an important indicator of shade-tolerance. ND12 had a rational allocation of assimilation to alleviate the negative effect on morphogenesis and physiological characteristics.
Illumina HiSeqTM2000 technique was used to sequence the Wuzhai Black Bean genes expression level infected by Heterodera glycines. A total of 20，980，831 reads with the length greater than 200bp have been obtained after splicing the two libraries， with the total length around 4.23Gb. 1045 differentially expressed genes were obtained. Gene ontology significant enrichment analysis displayed 1007 differentially expressed genes which enriched their biological processes. Orthologous genes classification in COG database showed that many differentially expressed genes were involved in carbohydrate and amino acid transport and metabolism， the synthesis of secondary metabolites, the signaling conduct mechanisms and the induced expression by H.glycines. KEGG metabolic pathways revealed differentially expressed genes with enriched phenylpropanoid and oxidative phosphorylation in metabolic pathways. This indicates these two metabolisms played an important role in resistance to H. glycines for Wuzhai Black Bean.
To investigate differentiation of Sclerotinia sclerotiorum pathogenicity, one S. sclerotiorum isolate was inoculated into rapeseed stem from cultivars with different resistance to the pathogen. After inoculation, 46 sclotia/strains from 46 inoculated rapeseed cultivars were collected followed by the second inoculation onto cultivar Qianyou 16 using 2 in vitro methods (toothpick inoculation and leaf inoculation). The second inoculation results showed that lesion length from these 46 sclotia/strains had significant difference. Both methods showed the same trends that the order of the isolate pathogenicity was mid-susceptive, high-susceptive, low-susceptive and resistant, according to the corresponding host cultivars. In other words, the isolate from resistant cultivar inoculum had the weakest pathogenicity, while the corresponding isolates from mid-susceptive cultivar inoculum had the highest pathogenicity. It suggested that the differentiation of S. sclerotiorum pathogenicity might be related to hosts’ resistance of oilseed rape.
Responses of seedling root to herbicide Attack were investigated under indoor hydroponics culture using 2 cultivars (Brassica napus cv Shaanyou 16 and B. rapa cv Baizayihao. Results showed that inhibitive effects of Attack on root were enhanced with increased herbicide concentrations. Main root length, root dry weight and root-shoot ratio were depressed. Root hair and lateral root lengths were shortened. Root hair density was reduced, root hair shape was distorted and their distributions were uneven. Moreover, MDA, POD and CAT activities of seedling roots were ascended with the increased herbicide concentrations. SOD activity increased first and dropped later. Attack swelled and thickened root tips. Root cap shape changed from taper to circular arc. High concentration of Attack led to disorganized array of epidermal cells and part of the cell missed. Cortical cell layers were lessened, cell vacuolation degree increased and cell edge was damaged. Results also showed that Baizayihao held stronger tolerance to Attack stress than that of Shaanyou 16.
This experiment was to investigate the inter-specific competitive interaction between Ageratina adenophora and Giycine max and to explore the methods of controlling the invasive weed A. adenophora displaced by G. max. In the field, soil tillage methods, cutting ways for A. adenophora and different density of G. max were considered. Plant height , density, competitiveness for light, nutrition and water of A. adenophora and G. max, and yield of G. max were investigated to evaluate the competition effects between A. adenophora and G. max. The results showed that A. adenophora was the dominant population and the growth and yield of G. max was inhibited when G. max seeds were sown between the plant gaps of A. adenophorum without tillage. G. max became the dominant species companioned with a significant competitive advantage of light usage when G. max seeds were sown after cutting A. adenophorum without tillage. Growth and yield of A. adenophorum were inhibited, and the absorbed nutrition and water were significantly reduced. With the increase of plant density of G. max, its yield increased significantly from 2113.66 kg/hm2 to 2885.62 kg/hm2.The invasive weed A. adenophora was displaced completely by G. max within rotary tillage treatment and the yield of G. max reached 3828.02-4007.87 kg/hm2. Through the suitable cultivation measures, the competitive advantage of G. max is very strong, and it can effectively replace A. adenophora, resulting in greater economic benefit.
Sample preparation and electrophoresis conditions play crucial roles in proteomic technologies, yet they can be problematic. In this study, we compared three protein extraction methods, including TCA/actone method, phenol extraction and phenol/SDS extraction. We also optimized the conditions and established parameters of two-dimensional electrophoresis for the soybean seed during germination. The results showed that phenol/SDS extraction was the most suitable one. For 2-DE, 0.2mg samples were applied into IPG strips of pH4-7(Bio-Rad,17cm), subjected to IEF as follows: 250V for 1h, 500V for 1h, 3000V for 3h, 8000V for 1h, 10000V for 1h, and then 10000V until reaching 60000Vh. This study established a solid foundation for the proteomic analysis of soybean seed germination.
Florisil extraction cartridge was tested to purify the enzymes-hydrolyzed oil followed by GC analysis. And the results showed that with Florisil extraction cartridge, sn-2 monoglycerol can be separated from the enzymes-hydrolyzed oil efficiently, and the mixture of triacylglycerol, diacylglycerol, monoglycerol and free fatty acid can also be efficiently separated in turn at the optimized extraction conditions. On this basis, 5 different type of oils (plant oil, microbial oil, algal oil, marine animals oil and fish oil) hydrolyzed by sn-1,3 specificity lipase were purified by Florisil extraction cartridge, then sn-2 monoglycerol were collected before methylating. Qualitative and quantitative analysis of sn-2 position fatty acid were performed by GC with a FFAP capillary column. The results showed that sn-2 position fatty acids in plant oil, algal oil and marine animals oil were mainly medium- and long-chain unsaturated fatty acids; sn-2 position fatty acids in microbial oil and fish oil were mainly saturated fatty acids, and in fish oils were mainly long chain polyunsaturated fatty acids.
An unknown bacterial strain was isolated from the edible natto and this strain could effectively degrade aflatoxins B1 (AFB1). The degradation rate of AFB1 could reach 91.4% with the concentrated crude enzyme solution prepared from the fermented supernatant. The strain was then taxonomic status identified and enzymatic properties of crude enzyme were investigated. Results were listed as follows: this strain was identified as Bacillus subtilis based on physiological, biochemical and 16S rDNA sequence analyses, thus named as B. subtilis sp. Natto3. The optimal growth time, reaction temperature, and pH value for the degradation of AFB1 by crude enzyme solution were 72h, 37℃ and 8.5 respectively; the degradation activity could be inhibited to different extents by 5 metal ions including Zn2+, Mn2+, Mg2+, Cu2+ and Li+. Furthermore, we added the crude enzyme solution into the peanut sample highly contaminated by AFB1, and examined whether AFB1 could be degraded. Detoxification experiment showed that AFB1 content in the contaminated peanut samples was decreased from 192μg/kg to 43μg/kg by the crude enzyme solution prepared from the culture of B. subtilis sp. Natto3.
To facilitate machinery harvesting of rapeseed, herbicide diquat was used as chemical ripener in field. The effects on dehydration and seed quality (including germination rate, thousand-seed weight, protein, fatty acids and diquat residues) were investigated using a Brassica napus cv Yangguang 2009 during mature season. Results showed that, compared to control (natural mature after water spray) and hand harvesting (swarfing), pods and seeds were remarkably dehydrated at 6d after 1.8 L/ha diquat treatment when the pods color on main inflorescence changed at ripening stage. The moisture level of pod and seed were 7.8% and 10.49% respectively, which were significantly declined by 52.66% and 24.84% respectively than that of control. The treated seeds germination rate, thousand-seed weight and quality had no significant change. Diquat residue in seed was 0.13mg/kg after 2 weeks in storage from harvesting, pass both Chinese and British testing standards. No residue was found after 3 weeks in storage from harvesting. In conclusion, it is necessary to promote diquat herbicide registration as ripener on rapeseed.
A synchronous detection method by HPLC-MS/MS of 5 pesticide residues in peanut was studied. After being extracted, cleaned-up synchronously with the distribution of liquid-liquid extraction method, acetochlor, isopropyl methyl grass amine, aldicarb, imidacloprid and phoxim in peanuts were analyzed by HPLC-MS/MS. The result showed that at the spiked sample levels of 0.01, 0.05 and 0.10 mg/kg, the average recovery rate was 80.9%~108.5% with the relative standard deviation of 1.5%~7.8% , and the detection limit was 0.001~0.011 mg/kg. This method was simple and economical, without requirement for special reagents and instruments. The detection sensitivity was high with the limits of detection and quantitation both below the national maximum residue limit. All indicators were conformed with the national quality control standards for pesticide residue analysis and this method could meet the requirement of routine analysis of pesticide residues in peanuts.
Rapeseed clubroot disease caused by Plasmodiophora brassicae is a serious disease in oilseed rape (Brassica napus L.) production areas in China in recent years. In this study, susceptive cultivar Zhongshuang 9 was used to test the protective effect of 15% Fluazinam. Results showed that this agent had no effect on seed germination, nor on seedling development, seedling height, root height and seedling fresh weight when the seeds treated with coating rate over 1:50, compared with those from non-treated seeds. Control efficiency to rapeseed clubroot were 43.7%, 46.4%, 83.5%, 92.5% in pot experiment, and were 22.2%, 53.6%, 44.5%, 54.5% in field experiment when using 15% Fluazinam suspension concentrate with seed coating ratio as 1:200, 1:100, 1:50 and 1:25 respectively. Thus the agent of 1:100 and 1:50 could be applied for clubroot management in oilseed rape production areas.
