KCS10 gene is one of the members of the β-ketoacyl-CoA synthase (β-ketoacyl coa synthase, KCS) gene family. To understand the function of KCS10 gene, genome walking and RACE techniques were used to clone BnKCS10 gene from B. napus cv. Zhongshuang 9. The BnKCS10 gene has a 2 344bp-coding region containing 2 introns spanning 527bp and 167bp respectively. BnKCS10 gene expressed at significantly higher levels in flower buds, and also in stigma and 30d developing seeds than in the vegetative organs. The yeast cells transformed with BnKCS10 gene produced KCS10 protein normally, but no synthesis of very long chain fatty acids was detected.
In order to find new plant antibacterial peptide, whole genome of Brassica napus was scanned and analyzed by means of bioinformatics. A gene of BnPCD842895 obtained as the preliminary candidate, which had similar sequence as the known antimicrobial peptides. The BnPCD842895 gene is 189bp long, encoding 48 amino acids. The complete gene was synthesized artificially through the overlap polymerase chain reaction (PCR), and was cloned into pET30a-EDDIE-GFP expression vector. The inclusion was expressed in Escherichia coli. After in vitro protein renaturation process and self-cleavage of fusion protein EDDIE, antibacterial peptide product was obtained without amino acid wastage. Bacteriostatic activity detection results showed that BnPCD842895 had strong bacteriostatic activity both for Gram positive bacteria and Gram negative bacteria.
To assess the distinctness of homonymous Brassica napus L. varieties from different resources, 42 named varieties in totally 91 rapeseed (Brassica napus L.) varieties were employed in this study. 29 characteristics used in DUS (distinctness, uniformity, stability) testing were examined by field trial in which homonymous varieties were grown side by side. At the same time, 43 SSR markers developed by our previous research were applied for fingerprint analysis. We evaluated the distinctness of homonymous varieties by characteristics used in DUS testing and SSR markers. Results showed that 39.3 percent (22 pairs) of homonymous varieties were greatly distinct from each other not only on morphological traits but also on SSR markers. 26.8 percent (15 pairs) of tested homonymous varieties were extremely similar and Kinship coefficient analysis revealed that these varieties have a very close relationship. The percentage of accession without significance difference was only 33.9 (19 pairs). These results suggested that it is a common phenomenon that homonymous varieties from different resources are actually homonyms. Therefore, it is suggested for department of varieties collection management to supervise homonymous varieties closely.
Eight different microsomal ω6 fatty acid desaturases and one plastidial ω6 fatty acid desaturase cDNA sequences, designated CtFAD2-1, CtFAD2-2, CtFAD2-3, CtFAD2-4, CtFAD2-5, CtFAD2-6, CtFAD2-7, CtFAD2-8 and CtFAD6 have been isolated from safflower (Carthamus tinctorius L.) by using PCR. All of the deduced amino acid sequences showed three histidine boxes characteristic of all membrane-bound desaturases, and CtFAD2 contained a C-terminal ER retrieval motif, while CtFAD6 possessed a putative N-terminal signal peptide. Our Blast searches of the deduced amino acid sequences revealed that CtFAD2-1 and CtFAD2-8 showed higher similarities to other plant seed-type microsomal ω6 fatty acid desaturases, whereas CtFAD2-2 showed higher similarities to constitutively–expressed type. The other safflower microsomal ω6 fatty acid desaturase genes showed much lower identities to plant FAD2. The CtFAD6 showed higher similarity with other plant plastidial ω6 fatty acid desaturases. Hydropathy analysis showed that the encoded polypeptide contains six putative membrane- spanning domains. Transmembrane analysis indicated that all the safflower fatty acid desaturases contained four to six putative membrane-spanning domains except CtFAD6, Protein second-structure indicated that safflower ω6 fatty acid desaturase is mainly composed of α-helix and β-sheet.
GmCAT3、GmCAT4、GmCAT5 gene were amplified by RT-PCR，Phylogenetic analysis indicated that GmCAT3 and GmCAT1/2, GmCAT5 and PsCAT, VrCAT had genetic relationship. The target genes were cloned into pMD19-T vector, and ligated to expression vector pET28a. The recombinant vectors pET28a/GmCAT3, pET28a/GmCAT4 and pET28a/GmCAT5 were transformed into E.coli BL21(DE3) and induced. The expression products were identified by SDS-PAGE. The optimal expression conditions of genes were 12 h under 20℃ by 0.1mmol/L IPTG inducing. They were expressed at 60kD molecular weight specific protein as inclusion body respectively. The proportion of recombinant proteins in total bacterial proteins were as follows: 35.7%, 44.2% and 42.5% under this specific conditions．
In order to establish an efficient peanut regeneration system for genetic transformation, embryonic leaflets of mature seeds of 7 varieties were used to develop regeneration seedling. The results showed that significant differences on frequency of adventitious bud induction and frequency of shoot formation existed in peanut genotypes, and Fenghua 2 was the best one among these cultivars. MSB + 6-BA 7mg/L + NAA 0.6mg/L + AgNO3 2mg/L was the best induction medium for Fenghua 2 adventitious shoot buds induction and shoot formation, with frequency of 57.22% for shoot formation. The suitable medium for buds regeneration and elongation was MSB + 6-BA 3mg/L; plantlets over 96% were rooting in MSB + NAA 1.5 mg/L or 1 mg/L. The regeneration plants above 90% were growing normally after being transplanted to soil.
Three chemical agents, GSC, SX-1 and Maiji, were applied for two years to research male sterilization on winter Brassica rapa cv Longyou 6 in northern cold regions. Results indicated that SX-1 and GSC had better inducing effects on stamens than Maiji. Concentrations of the chemical hybridizing agents (CHA), treatment period, spray times and dose had great influences on inducing effect. Tests showed that spraying twice led to the best sterilization effect. Double spray of SX-1 using 9.0 to 10.0mg/L with 3 to 4mL and 5 to 6mL per plant at budding stage and 10 to 12 days later, led to more than 95% plants being male sterile. Or, double spray of GSC using 0.6mg/L with 9 to 10mL and 15 to 16mL per plant, resulted in 98% plants being male sterile. When the dose was more than 8mL SX-1 or 20mL of GSC per plant, it would damage the plant.
Five high yield peanuts varieties, Xuhua 5, R03-3, Tangyou 4, Fenghua 5, and Shanyou 101 introduced from other regions of China were selected to study their podding characteristics in Liaoning Province. The results showed that number of gynophores, effective pods and well-filled pods per plant in Tangyou 4 and R03-3 were significantly higher than those in Xuhua 5 and Shanyou 101 (P?0.05). Ratios of gynophores number, total pods, effective pods and well-filled pods per plant at beginning of flowering stage to total flower number were the highest, followed at peak flowering stage. Few flowers could develop into gynophores at the end of flowering stage and none developed into pod. Rates of well-filled pod of R03-3 at initial and peak flowering stages (100% and 13.1%) were the highest. Tangyou4 and Shanyou 101 were followed, and Fenghua 5 were the lowest with 63.6% and 5.3% only. The highest pod positions were located on the 4-5th nodes of main stem in general. Much lower node position had greater number of pods. The highest pod position of Tangyou 4 was located on the 3th node of main stem, and that of Shanyou 101 was on the 6th node.
Net photosynthetic rate (Pn), stomatal conductance (Cond)，intercellular CO2 concentration (Ci) and transpiration rate(Tr) in leaves of sunflower at different development stages (two pairs of leaves, 13-14 leaves, bud stage, flowering stage, 20 days after blossom) were measured in an open-circuit gas channel system．Pn was simulated by Michaelis-Menten model，and then characteristic parameters of the light response curves were calculated．The results showed that Pn increased with increasing PPFD in 0 to 2 000 μmol•m-2•s-1 at each development stages, and the increased rate reached a maximum from vegetative to reproductive growth, then decreased with leaf senescence. During the whole growth period of edible sunflower in Hetao irrigation district, the apparent quantum efficiency mostly in the range of 0.067 to 0.083, the maximum net photosynthetic rate in the range of 14.61 to 64.16 μmol•m-2•s-1, light compensation point and light saturation point were 41.17 to 107.83 μmol•m-2•s-1and 2 382.89 to 3 600.51 μmol•m-2•s-1 respectively. It was shown that sunflower was the typical light-loving plant with high solar energy utilization efficiency. Pn, Cond and Tr increased significantly with increasing light intensity at different development stages of sunflower, while Ci reduced. When PPFD < 500 μmol•m-2•s-1 , WUE increased rapidly with increasing light intensity; when PPFD > 500 μmol•m-2•s-1, WUE still increased at two pairs of leaves stage, but changed as parabola at other developmental period.
Four isoenzymes of peanut lipoxygenases, B-Ⅱ，B-Ⅲ，C-Ⅴand C-Ⅵ were isolated form peanut seed by using ammonium sulfate precipitation, DEAE-52 ion exchange chromatography and Superdex G-150 gel filtration．These lipoxygenases were comprised of one equal subunit in SDS-PAGE experiment, and relative molecular mass of this subunit was 31kD. The final purification fold of B-Ⅱ, B-Ⅲ, C-Ⅴand C-Ⅵ were 63.2, 50.7, 22 and 55.45 respectively, and their yield of LOX were 4.8%, 5.3%, 2% and 10% respectively.
Surveys for stem canker/blackleg on oilseed rape (Brassica napus) were conducted from 60 locations throughout 16 provinces in China covering both winter and spring oilseed rape production regions during 2008 to 2012. Diseased plants were found in 42 locations from 14 provinces. One tenth of the plots were found to be blackleg-infested with up to 92% occurrence and 5% of plant death in the most severely affected plots. The disease symptom of plant was the same as that infested by Leptosphaeria biglobosa in Europe and North America. The morphological characteristics of ascospores released from pseudothecia were consistent with the description in the literature. The infected plants or plant debris with blackleg symptoms were collected and the causal fungus was isolated from the lesions. In the purified isolates, yellow to yellow brown or pink brown pigments were produced in PDA medium, which was the same to the control of L. biglobosa from Poland. Multiple PCR were used to identify 348 isolates out of 550 isolates and 468 diseased debris samples. A 440bp L. biglobosa production was detected. In vivo cotyledon inoculation results showed that all the 22 isolates tested from 11 provinces were pathogenic to B. napus and had higher pathogenicity than the control of Polish L. biglobosa isolate PL-Lb except CN-21from Hailer, Inner Mongolia. Among them, 14 were highly pathogenic and 8 were moderately pathogenic. The study indicated that blackleg caused by L. biglobosa had already existed in China and caused serious yield loss in some areas. Fortunately, no aggressive L. maculans was found. Thus a strategy to prevent L. maculans spread onto oilseed rape crops in China was suggested.
To improve the control techniques of rapeseed clubroot (caused by Plasmodiophora brassicae), the contolling effects of three fungicides were investigated during 2011-2012 under nursery bed and fields conditions respectively. Results showed that control efficiency was up to 90% by nursery bed disinfection with 0.5mL/L dilution of cyazofamid 2 to 3d before sowing. The disease occurrence of plants from cyazofamid disinfection combination with or without roots irrigation using chlorothalonil decreased 67.6% to 80.8% and 57.1% to 74.3% respectively, compared with non-treatment plants. The disease incidence of plants from seed treatment with fluazinam or cyazofamid decreased 75.4% to 95.3%, and 35.1% to 49.8% respectively, compared with plants from non-treatment seeds. The yield of plants from cyazofamid disinfection combination with or without roots irrigation by chlorothalonil increased 949.0kg/hm2 and 799.1kg/hm2, and the income increased 2 935.8 yuan/hm2 and 2 922.2 yuan/hm2 compared with non-control plants respectively.
To understand the synthesis of stilbene in different years and regions of peanuts after Aspergillus flavus infection, simple HPLC method was developed on simultaneous determination of six stilbenes (resveratrol, piceid, piceatannol, ε-viniferin, δ-viniferin, and pterostilbene) in peanut. HPLC separation was achieved on a Purospher STAR RP-18e column (Hiber150×4.6mm, 5μm) at 306nm, 30℃. A mobile phase of acetonitrile, water and 2% formic acid aqueous solution at a gradient elution with flow rate of 1mL/min was used. The method exhibited good linear relationships between the peak area and the concentrations range of 1 to 100 mg/L for the six stilbenes. The correlation coefficients were between 0.999 0 to 0.999 9. RSD of this method was less than 4.7%, the detection limits were between 0.01 and 0.05mg/L. The average recoveries were 95.2% to 102.0%. Four peanut varieties from different producing area and planting year were used to validate this method. Results indicated that the total amount of stilbenes varied slightly within the same producing area, with a value of 1.23 and 0.4μg/g respectively. However, significant differences were observed for the same varieties from different areas, possessing a value of 33.69 and 71.08μg/g respectively. A negative correlation relationship was also observed between the total amount of stilbenes and infection rate. The method is suitable for rapid and accurate determination of resveratrol, piceid, piceatannol, ε-viniferin, δ-viniferin, and pterostilbene from peanut seeds infected by Aspergillus flavus, and could be used for development of functional food and resistant breeding or varieties screening study of peanuts.
According to the optimization of equilibrium conditions and chromatography conditions, a method was developed to detect methanol, ethanol and isopropanol (IPA) used for soybean meal by headspace gas chromatography (HS-GC). By this method, residual solvent from 8 soybean meal samples were tested. Standard curve showed a good linearity range of this method. The detection limits of methanol, ethanol and IPA were 0.719, 0.420 and 0.214mg/L, the recovery rates were 101.1%, 99.4%, 97.8% respectively. A simple and convenient method was provided for the determination of residual solvents’ content in different kinds of soybean meal samples.
To improve the oil content of rapeseed, parent materials were used in breeding program shuttled from two sites at high and low altitudes during 2006 to 2011. One site was Dali in Shaanxi Province (E109.93, N34.80) at altitude of 368m. The other was Zhangye in Gansu Province (E100.45, N38.93) at altitude of 2 300m. The 62 parent varieties were divided into 4 groups of low, middle, high and extra-high oil content respectively. Ecological shuttle breeding effects showed that oil content of the same variety was improved greatly from 2006 to 2011. The average increase of oil content was 4.9% and 6.57% at Dali and Zhangye respectively. And the highest oil content reached 61.53% at Zhangye. Combining with elite agronomic traits breeding, high oil content (more than 50%) cultivar Qinyou 4 was released by this ecological shuttle breeding method.
To optimize hormone combination of soybean callus subculture, the hypocotyls segment of soybean seedling were used as explants, primary callus were induced. The methenatical model was established by quadratic orthogonal rotary composite experimental design and the concentration of hormones for callus subculture were optimized. Result showed than the determination conefficient R2 of the model was 0.916 1. Optimal combination of hormone for callus subculture of soybean was MS with 6-BA 0.5472 mg/L, NAA1.206 6 mg/L and 2,4-D 0.122 2mg/L.
Agronomic trait of five chufa varieties from different production areas were studied under plant mulched drip irrigation in Xinjiang. The results showed that round tuber type had the best adaptability，higher yield and oil content. Henan round variety showed particularly great tuber uniformity, ease of harvest; and had the highest yield and oil content which reached 17 513.25kg/hm2 and 26.79% respectively. The number of tillers and plant height of small tuber type were significantly higher than the others. Yield and oil content of large tube type were the lowest. Henan round tuber type was the most suitable for large scale cultivation in arid climate region of Xinjiang.
Abstract: Haploid material had great application value both in fundamental research and breeding programmes. The haploid could be produced by androgenesis (anther and microspore culture), gynogenesis (unfertilized ovules), intraspecific and interspecifc hybridizations. Anther culture method was the most researched and widely applied in flax (Linum usitatissimum L.) haploid breeding by far. Research of haploid breeding in flax was reviewed, and found that emphasis was on the factors involving anther culture, such as: genotype, pretreatment, the media and so on. The problem of haploid breeding in flax in our country was also discussed.
