To better understand the functions of dihydrolipoamide acetyltransferase (E2) gene in rapeseed (Brassica napus L.), CDS sequence of E2 gene from Brassica napus L. cv Zhongshuang 11 (designated as BnPLE2) was cloned and transformed into Arabidopsis thaliana. BnPLE2 full-length sequence (1 503 bp) included 6 exons encoding 500 amino acids with a predicted molecular mass of 25.17 kD and pI 9.04. The amino acids of BnPLE2 were highly conserved. Phylogenetic analysis indicated that BnPLE2 protein showed highest similarity with AtplE2 in Arabidopsis. Overexpression construct p35S::BnPLE2 was performed and transformed into Arabidopsis Columbia. The positive transgenic lines were obtained and the abnormal phenotype was observed, including inflorescence spike like cluster, compact flower arrangement, chrysanthemum shaped. In addition, the disappeared apical dominance of main inflorescence, increased lateral branch, pod deformity and stigma protrusion were also observed.
To develop cytologically stable disomic alien additions between Brassica napus (Huashuang 3) (2n=38, AACC) and Isatis indigotiga (2n=14, II), the cultivar Huashuang 3 was pollinated by one such monosomic addition carrying one specific chromosome of I. indigotiga, but with very low pollen fertility caused by male sterile cytoplasm. The disomic addition (2n=40, AACC+1IIg) with normal cytoplasm of B. napus was obtained by crossing Huashuang 3 with this alien chromosome. The cytological and genomic in situ hybridization (GISH) analyses for this addition showed the main normal chromosome pairing (20 bivalents) and segregation (20:20) during meiotic divisions of pollen mother cells, resulting in the high pollen fertility, seed-set and the progenies with alien chromosomes. But lagging chromosomes also occurred at low frequency during meiosis. This relatively stable disomic addition provided the novel material for genetic study of this medicinal plant as well as for Brassica breeding.
For high yield breeding under high density planting of rapeseed, effective silique number on main inflorescence was always expected. By using a new line of multi-silique on main inflorescence named 12R1402, 4 generations (P1, P2, F1 and F2) were derived from a cross 12R1402×Huyou 17. They were analyzed using mixed major gene plus polygene inheritance model to understand the inheritances of effective silique number on main inflorescence. Result showed that effective silique number on main inflorescence of 12R1402×Huyou 17 were dominated by 2 pairs of major genes with additive-dominance-epistasis effects plus polygene with additive-dominance effects (in E-1 model). The additive effects of major genes were 40.80 and 40.58, and the dominance effects of these 2 major genes were -32.62 and -0.75 respectively. Hereditability of major gene (σ2mg) in F2 was 60.38% and that of polygene (σ2pg) was 2.14%. It indicated that the effective silique number on main inflorescence in B. napus L. was dominated by major genes. Higher hereditability of major genes could be favorable to an efficient selection in early generations of effective silique number on main inflorescence breeding. In addition, environmental variance and genetic variance accounted for 37.48% and 62.52% of the phenotypic variance respectively, indicating that the environmental factors had great effects on effective silique number on main inflorescence in B. napus L.
To reveal sterility mechanism of a dominant thermo-sensitive genic male sterile line TE5A in Brassica napus, host proteins interacting with BnaA.tsMs protein were screened. Sterile flower buds (diameter ≤2mm) RNA from TE5A were sampled. Yeast two-hybrid (Y2H) cDNA library was constructed by SMART technique. Bait vector of pGKBT7-BnaA.tsMs was constructed and transformed into Y2H gold. Results showed that Y2H cDNA library of flower bud from TE5A was successfully constructed. The library capacity was about 2×107 cfu. The average cDNA length were more than 1.8 kb with a recombination frequency of 100%. Bait vector of pGKBT7-BnaA.tsMs was constructed. The plasmid had no virulence and self-activity to yeast. 49 interactive factors were obtained. Among them, 3 interactive proteins (BUBR1, cyclin-D3 and FBW2) were especially found interacting with BnaA.tsMs, which might regulate the fertility of TE5A.
The creation and analysis of interspecific hybrids between cultivated peanut and its wild relatives with interesting genes significantly improved genetic of peanut variety. F1 hybrid plants between a cultivar, Baisha 1016 (2n = 4x =40, AABB), and a diploid wild species A. macedoi (2n = 2x =20) was established through embryo rescue and verified by SSR markers, and characterized through observation of metaphase chromosomes stained with DAPI and meiotic behavior of chromosomes, and genome fluorescence in situ hybridization (GISH). The results showed that the F1 had 30 chromosomes which were equal to the sum of gamete chromosomes numbers of the two parents. A. macedoi showed bright centromere bands with DAPI staining and included a pair of “small chromosome. Some of A genome chromosomes of Baisha 1016 in F1 were observed to be hybridized with A. macedoi genome probe. The average chromosome configuration of F1 was 0.6 III+8.27 II+11.6 I, and chromosomes in meiosis anaphase I and meiosis telophase I showed unequal division. These results indicated the F1 was a real interspecific hybrid, and its wild parent A. macedoi might be an A genome species, indicating the high possibility for compensational translocation between A. macedoi chromosomes and A genome chromosomes of peanut in F1. F1 plants were highly sterile, which might be mainly resulted from the unequal chromosome division and non-tetrad division at anaphase and telophase of meiosis.
The zinc finger protein in plant plays an important role in resistance to abiotic stresses. In this research, a cold resistance gene, SCTF-1, was introduced into soybeans via pollen tube pathway transformation. Six positive transgenic plants were obtained by PCR detection. Southern blotting showed that SCTF-1 was integrated into the genome of recipient soybean as a single copy. The quantitative real-time PCR showed that the expression of introduced SCTF-1 gene increased significantly in root, stem and leaf compared to the non-transformed soybean. Under the stress of low temperature, compared to non-transformed plants, the relative electrical conductivity of transformed plants were significantly lower with 21.10% to 23.09% lower value; the content of malondialdehyde of transformed plants reduced 10.56% to 11.74%, the peroxidase activity of transformed plants increased 25.02% to 30.38%. There is no atrophy, wilting and rolling in the leaves of transgenic plants. Therefore, the cold tolerance of transformed plants was significantly enhanced by overexpressing of SCTF-1 gene.
To evaluate the genetic diversity and construct molecule identification of 124 soybean accessions introduced from Russia, 34 pairs of SSR primers distributed in 19 linkage groups were selected to amplify these germplasm, and 9 soybeans cultivars of Heilongjiang Province were used as control. The result showed that 140 alleles were detected from 133 samples. Numbers of alleles for each primer ranged from 3 to 10 with the average of 5.47, while polymorphic information content was 0.65. The 133 germplasm resources were divided into 2 clusters. The clusters had no significant correlation to the division of variety quality types. The band profile were converted to digits base on the fragment sizes and were analyzed by ID analysis 1.0 software. The result shows that the 133 soybean breed samples could be divided by only 7 primers named as Satt281, Satt373, Satt226, Satt590, Satt596, Satt259, Satt243, and thus a set of molecule ID of Russian soybean had been constructed.
to study the effects of calcium fertilizer on physiological characteristics of peanut at late growth stage on dryland, physiological characteristics, agronomic traits, pod yield and yield related traits of peanut variety Huayu 25 were investigated after application of calcium fertilizer in field experiments in 2014 and 2015. The results showed that different calcium fertilizer treatments significantly increased chlorophyll content, photosynthetic rate, NR activity, SOD, POD and CAT activities of leaves, root activity, and significantly reduced MDA content at pod-filling stage and maturation stage of peanut. Calcium fertilizer increased Y (II), ETR, qP and qL at pod-filling stage, while it decreased qN and NPQ. Calcium fertilizer could delay plant senescence of peanut, which were beneficial for peanut keeping their higher photosynthetic rate and longer photosynthetic time under drought conditions. Calcium fertilizer significantly increased the pod yield of peanut due to increasing pod number per plant, double kernel pod rate and seed plumpness. A rate of 14 kg/667m2 CaO was suggested to apply to peanut for achieving a high pod yield on dryland.
Thirteen soybean cultivars were used to evaluate the impact of maize-soybean relay strip intercropping on seed protein quality. The results showed that soybean protein content, relative content of lipoxygenase, total amino acids, and sulfur-containing amino acid contents were diverse among 13 cultivars in monocropping. Under intercropping condition, protein contents of majority cultivars had no significant difference between monocropping and intercropping except for 2 cultivars; Relative contents of lipoxygenase of 4 soybean cultivars decreased significantly; The changes of different protein subunits in different soybean materials and different protein subunits in the same soybean were not consistent; Sulfur-containing amino acid contents of 3 cultivars increased, and methionine contents of 5 cultivars increased significantly. The variable coefficients of histidine and arginine contents were the highest among soybean cultivars in intercropping. Clustering analysis employed was based on the changes in contents of crude protein, lipoxygenase, protein subunits and amino acids. The results showed that compared with monocropping, the protein content of Mengqing 6 and Caidou were not significant different, but relative percentage of 11S, content of total amino acids and sulfur-containing amino acid were increased, and percentage of lipoxygenase were decreased in intercropping. In conclusion, to meet different market demand of good soybean products, the selection of specialized soybean cultivars in combined with appropriate intercropping are essential in maintaining protein nutritional components, improving the flavor and taste of soybean products and extended storage period.
Large-seed peanut variety HY22 and small-seed peanut variety HY23 were used to investigate the effects of single-seed sowing with high, medium and low density on the activeness of key enzymes of carbohydrate and nitrogen metabolism in leaves, pod yield and kernel quality of peanut in field experiment. The results showed that carbohydrate and nitrogen metabolism level were significantly improved, activity of nitrate reductase (NR), glutamine synthase (GS), glutamate dehydrogenase (GDH), sucrose synthase (SS) and sucrose phosphate synthase (SPS) in leaves were significantly increased in single-seed sowing with medium and low density during the growing stage, while single-seed sowing treatment with high density had no evident difference compared with CK. For large-seed variety HY22 and small-seed variety HY23, under medium density in single-seed sowing, both of their pod yield were the highest with 8.1% and 7.3% increase respectively. The yield increase could be attributed to the higher pod number per plant, full-pod rate and higher pod weight. The kernel quality of large-seed variety HY22 were improved at medium and low density in single-seed sowing, while the kernel quality of small-seed variety HY23 showed no significant difference under the same condition.
Based on previous studies, predicting model on distribution of dry matter was constructed for castor. In the model, Richards function was used to express distribution index dynamic changes of leaves, stems, roots and capsule. The maximum distribution index was introduced to distinguish different genotype including Tongbi 5, Tongbi 6, Tongbi 9 and Zhongdong. The distribution index maximum value of leaves, stems, roots and capsule were given. The Nitrogen fertilizer influence factors was introduced to distinguish different levels of nutrient effects. Test results showed that absolute prediction error of dry matter weight was 0.64 - 19.34 g, and RMSE was 1.01 -11.43, which demonstrated that this model could provided an acceptable prediction.
The effects of tillage method with straw mulching on peanut yield and quality were studied by field experiment with split-plot randomized block design. The results indicated that peanut yield under ridge tillage combining with wheat straw mulching pattern was higher than that of no-tillage without or with wheat straw mulching patterns, and their yield increase were 19.4% and 25.0% respectively. Pod yields of cultivar Yuanza 9307 were higher than that of cultivar Yuanza 6 under 3 patterns with increase of 13.8%, 9.9% and 15.8% respectively. Peanut economical and agronomic characteristics showed the best results with the combination of ridge tillage and wheat straw mulching, including full pod number, 100-pod weight, kernel rate, and plant height, lateral branch length, branch number, valid branch number. These characteristics of Yuanza 9307 were better than that of Yuanza 6 under all 3 patterns. Tillage method and straw mulching had significant effect on NPK content in peanut kernels. Peanut protein and crude fat production with the combination of ridge tillage and wheat straw mulching were the highest, and peanut protein and crude fat production of Yuanza 9307 were greater than those Yuanza 6. Peanut yield, protein and crude fat production of Yuanza 9307 were the highest by ridge tillage with wheat straw mulching, and they were 4290.0 kg/hm2, 1294.3 kg/hm2, 1881.9 kg/hm2 respectively.
Rapeseed yield from main inflorescence under direct-seeding (high density) pattern makes great contribution to all plant yield. To promote main inflorescence yield, field experiment was conducted to research nitrogen (N) application effects, source-sink relationship and grain-filling process. The main infloresence of Brassica napus cv Huayouza 9 was divided into 3 parts (up, middle and bottom) according to 3 flowering periods (final flowering, nourishing flowering and early blooming). Results showed that N application improved seed yield significantly in these 3 parts. The corresponding increases were 73.3%, 138.3% and 108.6% respectively. Yield improvement was mainly resulted from more siliques in all 3 parts (53.4%, 37.9% and 25.2% more siliques respectively). 1 000-seed weight of middle part was improved, and seeds per silique on upper part was increased. On up and bottom parts, N treatment resulted in higher dry matters of silique shell and larger seed-to-shell ratio, which indicated that N accelerated material and energy transport from silique wall to seeds. N application also resulted in longer periods and more rapid dry matters and N accumulation in seeds. Duration of grain-fulling was important to silique numbers and seeds per silique. 1 000-seed weight was closely related to dry matter accumulation rate. Therefore, under direct-seeding high density pattern, N fertilization could improve source-sink relationship, accelerate grain-filling process, and further increase rapeseed yield.
Root-knot nematodes (RKN, Meloidogyne spp.) induce the formation of giant cell, namely root-knot, by directing the normal root cell of soybean plant into nematode-feeding cell. To investigate the role of hormone alternation in this process, contents of IAA, ABA, JA-Me, GA and ZR were monitored by enzyme-linked immunoassay method. Our results showed that contents of all these hormones were changed during the formation of giant cell in soybean roots. Among the hormones, the contents of JA-Me and ZR were consistently higher than those of control within 28 days after RKN inoculation. IAA content was significantly higher than that of control whereas ABA content was significantly lower than that of control within 5 days after inoculation. After that, IAA content was significantly lower than that of control, meanwhile contents of ABA and GA were consistent to those of control basically, except at 14th day and 21st day after inoculation, in which the contents of ABA and GA were higher than that of control respectively. Analysis of the five endogenous hormonal ratio changes showed that IAA/ABA and JA-Me/ABA ratio changed significantly, ZR/ABA and GA/ABA ratio were stable. These data indicated that IAA, ABA and JA-Me played an important role on information of root-knot during nematode infection. Our results also assisted in finding the physiological response mechanism of soybean responding to nematode infection, and to provide a new strategy for preventing and treating root knot nematodes.
European sunflower moth Homoeosoma nebulellum Hübner is the most serious pest on sunflower and seeds. To better understand the relationship between plant flowering phenology and host plant resistance to H. nebulellum, experiment was conducted to evaluate infestation index of seed from 12 typical sunflower accessions in field. Results showed that sunflowers during R5.1 to R5.2 flowering stages (opened tubular florets accounting for 10-20% of whole head) were damaged more heavily than those at other flowering stages. The reason could be the coincidental occurring R5.1-R5.2 flowering stage and the peak of adults moth emergence and oviposition. Oilseed material MGS identified with higher resistant to sunflower moth, conversely showed higher infestation index of seed than others, as its flowering stage was in R5.2 flowering stage. On the contrary, confection cultivar Xinghuo identified with higher susceptibility, exhibited the lowest infestation index of seed due to its delayed flowering stage. Therefore, planting time should be take into consideration to avoid the occurrence of adults moth and to alleviate its damage.
to optimize nutrients quantitative analysis in edible oil, we developed an LC-MS/MS method coupled with ultrasonic extraction to analyze phytosterol and tocopherol. Central composite design was employed to optimize the extraction conditions including ultrasonic time and ultrasonic temperature. After systemic analysis, the optimized experimental conditions were obtained as: ultrasonic temperature at 75℃, ultrasonic time of 40 min and ultrasonic power of 100%. Validation indicated that the limits of detection were 10.0 ng/mL, 10.0 ng/mL and 8.1 ng/mL for stigmasterol, β-sitosterol and δ-tocopherol respectively. Recoveries ranged from 87.2% to 97.8%. Correlation coefficients were higher than 0.990 4. And the relative standard deviations were 2.3% to 8.6%. Consequently the method was used to analyze stigmasterol, β-sitosterol and δ-tocopherol in camellia oil, perilla oil, olive oil and soybean oil. Results showed that this method was simple, rapid, accurate and reliable, suitable for quantitative analysis of main phytosterols and tocopherols in vegetable oil.
To investigate changes of pressed oil after microwave pretreatment of seeds, hemp seeds were treated with microwaves under 560 W for 7 min at a frequency of 2 450 MHz. After the pretreatment, seed oil was extracted by cold press. Quality and antioxidant activity of oil were consequently measured. Results showed that microwave significantly increased acid values and peroxide values, and decreased moisture content of oil. The color of oil was deeper than that of untreated. Dehulling significantly improved oil color. Fatty acids were detected in oil including 13.88% oleic acid, 57.59% linoleic acid, and 16.33% linolenic acid. 3.53:1 ratio of n-6 and n-3 fatty acids was found in oil which was a well-balanced proportion of the 2 essential fatty acids. Microwave had no significant effect on fatty acid composition in oil. Total phenolic content was increased by one fold, total tocopherols and phytosterols contents were increased by 41.99 mg/kg and 58.68 mg/100 g respectively. Microwave significantly improved oxidative stability of oil and its antioxidant activity. The induction period (IP) of oil was increased 1.05 h, DPPH and FRAP values were increased 8.46 and 81.85 µmol/100 g respectively. In conclusion, it is recommended to treat hemp seeds by microwaves before cold press for oil.
To explore optimized factors on sporulation of Colletotrichum truncatum, effects of different media, illumination, temperature, and sucrose concentrations on conidial production and the effect of sucrose concentration on pathogenicity were studied. The results showed that optimum medium was oat medium among 4 tested media. Illumination and temperature had no obvious effect on sporulation. Conidial production reached the maximum on the medium contained 9 g/L sucrose. Re-inoculation experiment indicated that sucrose concentration had no significant impact on pathogenicity. In conclusion, Media type and sucrose concentration had significant effect on conidial production of Colletotrichum truncatum.
To screen effective fungicides for sunflower rust caused by Puccinia helianthi, 15 fungicides were tested by measuring toxicity on urediniospores. Results showed that urediniospores of the pathogen were most sensitive to 20% sodium dichloroisocyanurate wettable powder (WP) with EC50 of 4.959 1μg/mL, followed by 10% difenoconazole water dispersible granules (WG) and 40% flusilazole emulsifiable concentrate (EC). To optimize effective spray time, fungicides effects were tested by spraying the fungicides after or before inoculating urediniospores on young leaves under green house conditions. Timing tests results showed that control effects of spraying before or after the inoculation were both sufficient. The most effective fungicides were 80% tebuconazole (WG), 25% triadimefon suspension concentrate (SC) and 20% sodium dichloroisocyanurate (WP) with 100% control efficiency which could be chosen to control sunflower rust in the field. 75% Chlorothalonil (WP) and 77% copper calcium wettable powder (WP) were not as effective in controlling the rust but could be applied as preventive chemicals.
