Diploid wild Arachis species A. stenosperma is an important genetic resource for the improvement of resistances to root-knot nematode, rust and late leaf spot in cultivated peanut varieties. In this study, a new interspecific hybrid, w1401, was developed by crossing cultivated peanut variety Yuhua 15 with the diploid A. stenosperma, followed by subsequent embryo rescue. Then, w1401 was characterized using Transposable Elements marker and sequential fluorescence in situ hybridization (FISH) with probes of telomere repeats, 5S rDNA, 45S rDNA and total genomic DNA of A. duranensis, A. stenosperma and A. ipaënsis. The result indicated that, the molecular and karyotype traits of w1401 three genomes were highly identical to the A and B genome of Yuhua15 and A. stenosperma genome respectively. Moreover, 34 specific SSR and transposable elements markers were developed, 32 of which were co-dominant markers with Yuhua 15, and could be used to identify translocation or introgression lines with A. stenosperma chromosome fragment in future studies. In addition, the differences between w1401 and its parents Yuhua 15 in phenotypic traits and resistance were also analyzed, w1401 showed better resistance to late leaf spot than that of Yuhua 15, indicating a promsing breeding future.
Multi-parent advanced generation inter-cross (MAGIC) population became a new tool for genetic mapping and breeding in crops. To explore the potential application of MAGIC lines on rapeseed breeding, a population encompassing 680 lines was developed by crossing 8 elite parents. All lines advanced by single seed descent method. At F3 generation, several phenotypes including disease resistance, yield potential and plant architecture were investigated, and 204 of 680 lines were chosen to form a sub-population for further selection. At F6 generation, several main agronomic traits of 204 lines and 8 parents were collected. The traits included plant height, number of primary branch, effective length of main raceme, effective siliques of main raceme, silique length, seeds per silique, thousand seeds weight and plant yield. Results showed that variation of main agronomic traits in this population had normal distribution. Differences of 5 traits were significant comparing to their parental population. The 5 traits included number of primary branch, effective length of main raceme, silique length, seeds per silique and thousand seeds weight. It indicated that MAGIC population was the ideal substitute for elite lines selection by traditional method in rapeseed breeding. Seed yield and ecological adaptability were tested in 3 sites (Jingzhou, Gong’an, Yuxi) for 2 years. 9 elite lines were obtained with better resistance to Sclerotinia sclerotiorum, high yield and high quality performance.
In order to research the biological functions of SGF14h and provide theoretical basis for studying the expression regulation of SGF14h, which was screened from pre mass spectrometry data in our lab, the bioinformatic computer software, database and online programs were used to predict and analyze the nucleotide sequence and amino acid sequence of SGF14h. The results of the gene structure analysis showed that SGF14h was located on the 4th soybean chromosome and its full length was 1788 bp including 780 bp ORF which encoded 259 amino acids. The results of protein properties analysis showed that SGF14h sequence was rich in phosphorylation sites, including 10 serine phosphorylation sites and 3 threonine phosphorylation sites; SGF14h had no signal peptide and no transmembrane domain, so it belonged to non-secretion protein; it was found that SGF14h was located in mitochondria by subcellular localization prediction. The results of the protein structure analysis showed that there were 153 α-helix, 0 β-sheet plus and 0 β-turn in SGF14h; SGF14h was a kind of hydrophilic protein and belonged to 14-3-3 protein family. The results of phylogenetic tree analysis showed that SGF14h had the closest relationship with GF14 protein of Maize. In dwarf soybean, SGF14 possibly played a key role in GA signal pathway and then affected the plant height. Therefore the study on SGF14h had biological significance, and these results could be the theoretical basis and provide evidences for further verifying the biological functions of SGF14h.
To meet the future demand of winter rapeseed identification, a DNA fingerprint database containing genotypic data of 419 new winter rapeseed varieties (lines) × 38 markers were constructed by capillary electrophoresis detection method with fluorescent SSR markers. 41 locus and 129 alleles were detected by 38 fluorescent SSR primers in 419 newly bred winter rapeseed varieties (lines) . Among the 38 markers, 35 were for 1 locus and 3 for 2 loci. The number of alleles per loci ranged from 2 and 6 with an average of 3.15 per locus. The number of genotype were 250 and varied from 3 to 15, with an average of 6.09 per locus. The polymorphic information content values ranged from 0.1322 to 0.6907 with an average of 0.39. Missing statistics calculated fingerprint database at the rate of 1.1%. 87571 data items were obtained by comparing DICE similarity coefficient among varieties (lines). DICE similarity coefficient among varieties (lines) ranged from 0.366 to 0.983 with an average of 0.692. Within the DICE similarity coefficient range from 0.601 to 0.800, there were 78571 data items, which accounted for 89.72% of the total. The results showed that the genetic of winter rapeseed varieties (lines) had limited diversity.
In order to assess the genetic diversity of peanut in Jiangsu Province, 133 accessions of peanut landraces were evaluated by 25 SSR markers. 93 alleles were amplified, and the average number of alleles per locus was 3.72. The polymorphic information content (PIC) of each primer varied from 0.066 to 0.749 with an average of 0.374. Shannon?s information index of each primer varied from 0.161 to 1.385 with an average of 0.671.The similarity coefficients of 133 peanut landraces varied from 0.416 to 1.000 with an average of 0.830. The order of average similarity coefficient of 4 types of peanut was Arachis hypogaea L. var. hypogaea (0.899)> var. hirsute (0.776)> var. vulgaris (0.710)> var. fastigiata (0.700). Cluster analysis with UPGMA showed that the landraces could be divided into three groups at 0.600 of genetic similarities coefficient. The results showed that there were rich genetic diversity of peanut landraces in Jiangsu province, and the genetic diversity was significantly lower among the var. hypogaea than those of the other three types. The result of cluster analysis had important significance for efficiently using these peanut landraces.
The correlation analysis of yield of progeny lines derived from cultivated soybean × wild soybean and their agronomic characteristics were performed to assess these lines and their agronomic characteristics affecting soybean yield. 20 agronomic characteristics of 18 stable progeny lines were analyzed by factor analysis. The quantitative trait analysis of plant and seed of progeny lines showed that the agronomic characteristics of each line had significant difference. 5 common factors were extracted by factor analysis, and the cumulative variance contribution rate was 73.611%. This result indicated these 5 common factors could primely reflect the information of these 20 traits, and the factor analysis was reliable for assessing the effect of agronomic characteristics on yield. Integrated assessment of each line and its agronomic characteristic, was used to screen out 5 strains of promotional value. The results showed that agronomic characteristics such as three-pod number, first-class branch, stem weight and effective branching could be used for evaluating soybean yield. 5 lines including 15, 3-2-2, 60Co 600, 18-2 and 8-1-2 were selected for further promotion, which could help to utilize and protect wild soybean resources.
To better understand the relationship between cold resistance and ABA content in winter rapeseed (Brassica rapa L.), the changes of ABA was measured using 2 ultrastrong cold-resistant cultivars (Longyou 6 and Longyou 7) and 2 weak-resistant cultivars (Tianyou 2 and Tianyou 4). Results showed that except Tianyou 4, ABA content increased gradually with the temperature before winter. Ultrastrong cold-resistant cultivars (Longyou 6 and Longyou 7) increased earlier than weak-resistant cultivars Tianyou 2. Correlation analysis indicated that wintering rate had greater correlation with ABA content of root than that of leaf. The ABA content of elongation part of leaf was higher than other parts in the mean time.
Soil alkaline stress is one of the main environmental factors affecting plant growth. Peanut (Arachis hypogaea), an important oil and economic crop in China, has been deemed to be with alkaline stress resistance. However, its response mechanisms to soil alkaline stress are still unclear. A pot experiment was used to investigate the effects of soil pH on root morphology and growth development characteristics at seeding stage of two peanut varieties. The soil pH were 8.5, 9.0, 10.0 and 6.0 (CK). The tested large seed variety Huayu 22 and small seed variety Huayu 20 were two main popularized varieties in Shandong province, China. The results showed that, (1) the inhibition of growing development of peanut were relatively small in pH8.5 treatment. Only root length and plant dry weight of peanut in pH8.5 soil were lower than those of CK. (2) Inhibition degree of peanut roots increased with the increasing of soil pH. In pH9.0 treatment, total root length, root volume and root surface area were reduced by 73.3%, 61.3% and 65.5% for Huayu 20 and reduced by 69.6%, 55.1% and 63.9% for Huayu 22, when compared with CK respectively. In pH10.0 treatment, the 3 indexes mentioned above were 98.9%, 81.1% and 96.6% lower for Huayu 20 and 98.8%, 85.1% and 96.8% lower for Huayu 22 than CK. The ratio of root length, root volume and root surface area with diameter 0-1mm to total root decreased and the ratio with diameter ≥2mm increased in pH9.0 and pH10.0 treatments. (3) Alkaline stress of high pH inhibited above ground parts of peanut seedling. Leaf area, dry weight of plant organs and root shoot ratio were decreased significantly in pH9.0 and pH10.0 treatment. Dry weight per plant in pH9.0 treatment was decreased by 43.0% for Huayu 20 and 29.4% for Huayu 22 compared with CK. In pH10.0 treatment, dry weight per plant was decreased by 74.8% for Huayu 20 and 69.7% for Huayu 22 compared with CK. Taken into consideration of all characteristics, soil pH exceeded 8.5 was not suitable for planting peanut. The peanut variety Huayu 22 had a greater alkaline tolerance than Huayu20.
To evaluate the protein nutrient of forage rapeseed, Brassica napus cv Huyouza 62 was studied based on WHO/FAO criteria model about essential amino acid (EAA). The essential amino acids for nutritional values were measured on Huyouza 62 protein at different growth periods. Results showed that rapeseed protein began to decrease gradually with the growth periods, but amino acids were rich in different stages. The content of essential amino acid in the flowering stage were Met+Cys (3.36%), Phe+Tyr (10.11%) and Ile (5.64%), Leu (9.45%), Val (6.73%) in initial podding, which were higher than those of other growth periods of the total amino acids. In flowering and initial podding stages, ratio of total essential amino acids to total amino acids (TEAA/TAA) were 44.88% and 44.36% respectively. The ratio of total essential amino acids to total non essential amino acids were 0.81 and 0.80 respectively, which were higher than those of other periods. By the amino acid ratio coefficient method, the essential amino acid index in initial podding and podding stages were much higher than that of other periods. The score of ratio coefficient (SRC) of amino acid was up to 82 at the initial podding stage. According to the feed criteria for cattle, sheep and goat, all the essential amino acids index of Huyouza 62 was greater than 0.95. When combined with biological yield and forage palatability, it could be concluded that Huyouza 62 plants at flowering and initial podding stages could be developed as new feed for ruminant animals.
To explore the effects of nitrogen form and ratio (NH4+-N/NO3--N ratio) on growth and nutrient uptake of sesame seedlings, 2 sesame cultivars Zhongzhi 13 and Luo 12 were employed in hydroponics with 7 NH4+-N/NO3--N ratio treatments (10:0, 9:1, 3:1, 1:1, 1:3, 1:9, 0:10. pH values of nutrient solution and plant phenotype were regularly measured. Plant biomass, root morphological parameters, and N, P, K, Ca, Mg, free NH4+-N and NO3--N concentration of leaf and root were determined at the 28th day treatments. Results showed that high NH4+ /NO3- ratios (10:0, 9:1, 3:1) inhibited seedling growth, and the inhibition increased with NH4+-N. Cultivar Zhongzhi 13 showed more tolerance to NH4+ toxicity. Under NH4+-N/NO3--N ratio at 1:9, seedlings exhibited the greatest plant biomass, and pH of nutrient solution maintained suitable especially for Zhongzhi 13. Compared with total NO3--N treatment (0:10), plant dry weigh, total root length and root surface area of Zhongzhi 13 were significantly increased 47%, 29% and 15% respectively. Moreover, appropriate NH4+-N/NO3--N ratio promoted the absorption and utilization of N, P, K in Zhongzhi 13, as indicated by N, P, K concentrations increase in leaves and roots respectively. However, compared with total NO3--N treatments (0:10), ratio 1:9 was appropriate with no effect on plant biomass and nutrients uptake of Luo 12. Therefore, appropriate NH4+-N could enhance sesame seedlings biomass and root growth, NH4+-N/NO3--N ratio of 1:9 was more suitable for growth and nutrient uptake of the seedling. It was recommended that NO3--N could be used as a major N source for sesame, and suitable NH4+-N for specific sesame cultivars and soil was suggested in field.
Pathogenesis-related proteins were originally discovered to accumulate in plants that were infected by pathogens，but were later found to accumulate in plants in response to a variety of biotic and abiotic stresses．It played a very important role in the process of plant disease. In order to investigate the expression of pathogenesis-related proteins (GmPRs) in soybean roots after inoculated by Heterodera glycines. Six GmPRs genes expression were examined after inoculated by Heterodera glycines in resistant cultivar Huipizhi heidou and susceptible cultivar Liaodou 15 by using real-time quantitative PCR (RT-PCR) technology and subcellular localization. Furthermore expression vector was constructed to research the subcellular localization of GmPR2 and GmPR10.The results showed that, in the resistant cultivar，the expression of GmPR2（β-1,3- glucanase）were significantly induced by inoculation treatment at early stage of inoculation and development.The difference of expression was not significant in susceptible cultivars, indicating that GmPR2 may also play an important role in soybean against Heterodera glycines;GmPR3 and GmPR10 reach the peak at the stage of development indicating that it could be related to prevent nematode development.Through subcellular localization, GmPR2 mainly located in cytomembrane, GmPR10 mainly distributed in the cytoplasm and nucleus, which was consistent with the predicted results. The transcript levels of GmPRs gene increased in different degrees after inoculated by Heterodera glycines, the expression of GmPRs proved that it played a different role in the defense of soybean against soybean cyst nematode.
Cry8-like gene was recombined into pCAMBIA3301 vector and a root-specific plant expression vector, and pCAMBIA3301-RSP-Cry8-like-nos with glufosinate resistant marker was constructed successfully by Seamless Assembly Cloning. Two different soybean varieties ?Jinong 28? and ?Jinong 42? were transformed by Agrobacterium-mediated genetic transformation. Detection of PCR showed that 17 transformed plants in T1 generation of Jinong 28 and 13 of Jinong 42 were obtained, and 36 transformed plants in T2 generation of Jinoing 28 and 25 of Jinong 42 were then produced. Southern blotting of T2 generation indicated that target gene was integrated into the genome of oybean with single copy. Detection of the transcription of Cry8-like by using quantitative real-time PCR (qRT-PCR) showed that Cry8-like expressed in root of transformed plants, while fluorescence signal was not detected in non-transformed control. The results of insect resistance to Holotrichia parallela larvae in laboratory showed that the ability of insect resistance was improved in transgenic plants.
In recent years, sclerotia mycelial germination (caused Sclerotinia stem rot) was found at oilseed rape seedling stage in Sichuan and Hubei provinces. To better understand this new type of sclerotinia rot, studies were carried out in relation to myceliogenic germination. Rapid water uptake was observed in sclerotia during the first hour and sclerotia were saturated in approximately 2 h. Saturated sclerotia had doubled in weight and geminated myceliogenically within 4 days of incubation without exogenous nutrients at 25 oC. Almost all sclerotium germinated myceliogenically within 11 d of incubation. High soil moisture was favorable for sclerotia to germinate myceliogenically. Younger sclerotia germinated faster. But after 9 d, the difference of germinating speed was not significant. Sclerotia could germinate myceliogenically at 10.0-30.0 oC, and germinated rapidly at 20.0-27.5 oC. The germination rate of sclerotia decreased with pH increasing. The highest germination rate was observed at pH 3 on the 4th day. Field inoculation test showed that sclerotia germinated myceliogenically. The mycelial germination caused Sclerotinia stem rot before initial flowering stage. The highest disease index reached 11.7% on rapeseed seedling.
Six biocontrol actinomycetes strain against Heterodera glycines were isolated from soybean rhizosphere at Heihe, Heilongjiang Province. To evaluate the field control efficacy of these strains, soybean seeds coated with fermentation broth of 6 strains were sown in disease nurseries at Heihe and Daqing respectively. The results showed that 6 biocontrol strains had significant resistance to first generation cyst nematodes production at soybean seedling stage. XFS-4 had the highest suppression rate, H. glycines of the soil around per soybean plant root was suppressed by 40.06% and 59.77% in Heihe and Daqing respectively. Identification based on its morphology, physiology and biochemistry and its 16S rDNA sequence analysis showed that the strain was Streptomyces zaomyceticus.
To evaluate the effects of Monolepta hieroglyphica (Motschulsky) adults on the damage to soybean yield loss and the economic threshold of this pest, the agronomic yield characteristics of soybean influenced by M. hieroglyphica in different densities were tested with the method of systemic field investigation and plot grid connection. The results showed that the hazard summit period of M. hieroglyphica adults was during period of July 26 to August 22 in soybean fields, a total of 40 days. During this period, some agronomic yield characteristics of soybean changed, such as: reduced effective pod number, increased rate of empty pod, decreased 100 kernel weight, and the significantly decreased yield of soybean. The relationship of population density of M. hieroglyphica adult and rate of empty pod fitted well to the logistic growth model y=8.720/(1+e(0.7189-0.026 0x)). Equally, a quadratic curve model y=19.6085-0.155x+0.0024x2 indicated the relationship of population density of M. hieroglyphica adult and 100 kernel weight, and a modified exponential model y=53.2078+33.4089×0.9408x(p<0.0001) for soybean yield, a Quadratic function model y=-0.6028+1.8126x-0.0258x2 (p<0.01) for rate of yield loss. Economic loss were 2.81% and 2.91%, and economic threshold were 193.63 bugs / 100 plants and 199.47 bugs / 100 plants, when lambda-cyhalothrin and methomyl were used for controling M. hieroglyphica in present soybean production.
A procedure for ultrasound-microwave assisted extraction of isoflavone from sprouts of Kennong 4 soybean was optimized using the orthogonal test. The particle size distribution of sprout samples after extraction was determined by laser diffraction particle sizing method. The antibacterial activities of isoflavone extract against Escherichia coli and Staphylococcus aureus were analyzed by broth dilution method and Oxford cup method. The results showed that the optimal extraction condition included ethanol concentration of 50% (v/v), material-liquor ration 1:25 (g·mL-1), extraction temperature 65℃, and extraction duration 250s. The best extraction rate of 0.387 was achieved under this condition, and particle size of soybean sprout samples from ultrasound-microwave assisted extraction was significant increased than that from heating extraction, which indicated that the loosened structure and increased volume of samples after extraction by microwave-ultrasound synergism could facilitate the mass transfer between isoflavone and extraction. Isoflavone extract had significant antibacterial effect against Staphylococcus aureus , and the minimum inhibitory concentration (MIC) was 265 mg·mL-1, however, had little effect on Escherichia coli.
To determine the contents of lignans and its components in flaxseed with different cultivars in China, 24 cultivars of flaxseed samples were collected from the main producing areas (including Inner Mongolia, Gansu, Shanxi, Hebei, Ningxia, Xinjiang), and the content of lignan and its components were determined by HPLC. The results showed that the alkaline hydrolysates of flaxseed lignans were p-coumaric acid glycoside and SDG; and the acid hydrolysates were SECO, p-coumaric acid and HMF. SDG was the highest in flaxseed lignans. The content of lignans hydrolyzate of flaxseed in different cultivars and areas was significantly different (P <0.05). Among 24 cultivars of flaxseed samples, the contents of CouAG (6.07 ± 1.97mg CouA/g), HMF (33.31 ± 0.50 mg/g), SECO (5.03 ± 0.16 mg/g ) and CouA (0.87 ± 0.01 mg/g) in Baya 9 were the highest, the contents of CouAG (1.16 ± 0.15 mg CouA/g ), SDG (11.37 ± 0.77 mg / g ) and CouA (0.18 ± 0.01mg/g) in Dingya 23 were the lowest. The content of SECO in Neiya 9 was lower than 1 mg/g seed (0.76 ± 0.01 mg/g ), and the content of HMF in Lunxuan 2 was the lowest at 0.13 mg / g . The content of SDG and its hydrolysates of flaxseed in Inner Mongolia, Shanxi and Hebei was relatively higher among the 6 flaxseed producing areas.
Lipid exerts numerous functions in plant growth and development such as components of biological membranes, carbon energy storage, protective tissue formation and resistance of pathogen infection. Lipid transport plays an important role in plant lipid metabolism. Plant lipids synthesis occurs in plastid and endoplasmic reticulum, where the generated lipid metabolic intermediates will be distributed to other compartments or out of the cells with different transport, via transporters, membrane contact sites, vesicular trafficking, or diffusion for lipid homeostasis. In this review, we summarized the recent research progress of the regulation mechanism of lipid transport among endoplasmic reticulum, mitochondria, peroxisome, or out of cells via diverse lipid transporters.
Rapeseeds are a rich source of phenolic compounds. The predominant phenolics present in rapeseeds are phenolic acids and condensed tannins. Among them, phenolic acids are present in the free, esterified and insoluble-bound forms and are derivatives of benzoic and cinnamic acids. Free phenolic acids account for 9-16% of the total, with sinapic acid being the most important compound of this group (over 70% of the total free phenolic acids). The largest fraction of polyphenolic compounds, accounting for about 80% of the total amount, corresponds to esterified phenolic acids, where sinapine, the choline-ester of sinapic acid, is by large the most representative one. Rapeseed phenolic were extracted with different systems. Extraction with organic solvent, special 70% methanol, is one of the most commonly used. Moreover, ultrasonic assisted extraction can improve effectiveness. Folin-Ciocalteus method, vanillin method, and HPLC analysis usually are used to measure total phenolics, condensed tannins, and phenolic acids content. Rapeseed phenolic extracts show excellent antioxidant activity, which usually were evaluated by DPPH, FRAP, and ORAC methods. The chemical structures of phenolics play key roles in antioxidant activity, depending on the number and position of free hydroxyl and methoxyl groups attached to the aromatic ring. The derivatives of cinnamic acid show a higher correlation with antioxidant capacity than the derivatives of benzoic acid. Heat treatment of rapeseeds can significantly increase phenolics and canolol content in rapeseeds and oil from them. Canolol can be formed by thermal decarboxylation of sinapic acid occurring naturally in rapeseeds. Approximately 56-83% of canolol are transferred to the oil after pressing, due to the strong fat solubility of canolol. Phenolic compounds and canolol are practically completely removed from oil during neutralization, bleaching, and deodorization steps. Therefore, unknown phenolic compounds could be identified by HPLC-MS/MS and NMR. The antioxidant activity of individual phenolic compound could be evaluated by cellular antioxidant activity assay, to provide a basis for the development and utilization of rapeseed polyphenols. Moreover, the moderate refining technology of oil could be developed to reduce the loss of polyphenols in rapeseed oil.
