For high yield breeding and light transmission improvement in field, 2 lobbed-leaf-margin mutants ll1 (interspecific hybridization between Brassica napus and B. juncea) and ll2 (selected from cross of 2 B. napus lines) were studied on their inheritance and agronomic traits. Classical genetic study showed that ll1 was controlled by one pair of recessive genes (previous study), and ll2 was controlled by another pair of dominant genes. Populations of F1, F2, BC1 and RBC1 (from reciprocal crosses) were obtained from the mutants’ hybrids. Inheritance results showed that the genes in ll2 were dominant over those in ll1. Their genes in ll1 and ll2 were closely linked. The leaf traits indicated that leaf areas of mutant seedlings were both significantly less than those of their wild types before winter. The yield traits (including primary branch, siliques per plant, seed per silique and seed yield per plant) of both mutants made no significance to the final yield. It implied that lobbed leaf margins had no negative effects on yields. These studies offered better understanding of ideal plant type and high yield breeding in rapeseed.
To develop Brassica napus germplasm resources for mechanization breeding, hybrid progenies from B. napus and Sinapis alba were analyzed on silique shattering resistance, comparing with B. napus cv Yangyou 6. Random impact test demonstrated that silique shattering resistance index ranged from 0.14 to 0.99. The frequency distributions among the materials were 6, 2, 4, 2, 5, 6, 4 and 1, when the silique shattering resistance index was divided into 0.1-0.2, 0.2-0.3, 0.3-0.4, 0.4-0.5, 0.5-0.6, 0.6-0.7, 0.8-0.9 and 0.9-1 regions, indicating wide variation in shattering resistance. Twelve lines demonstrated better resistance than Yangyou 6. Based on correlation analysis, the silique shattering resistance index had significant positive correlations with silique length, silique wall weight and thickness. Therefore, the long silique，thick and heavy silique wall could be used as auxiliary index to screen the silique-shatter-resistant rapeseed.
In this study, we characterized a set of 46 accessions of soybean from Russian Far East and 44 accessions from Heilongjiang Province using 40 SSR primers to evaluate their genetic diversity and population structure. The results showed that the genetic diversity of soybeans from Russian Far East was higher than that of Heilongjiang Province. The germplasm resources were divided into 3 clusters by NJ phenogram depicting and Structure software. Each cluster contained accessions from Russia and Heilongjiang Province, but the majority were likely originated from the same region. The majority of clusterⅠ, clusterⅡ were the accessions from Russia, and cluster Ⅲ were from Heilongjiang Province. In the present study, these germplasm resources were not divided in accordance with their geographical origin. It indicated that the relationship between soybean germplasm from Russian and Heilongjiang Province was close and it was necessary to introduce a wider range of germplasm from Russian Far East to broaden the genetic basis of soybean in Heilongjiang Province.
Key words: soybean; germplasm resources; genetic diversity; population structure
To breed high oleate peanut cultivars suitable for Shandong and Huang-Huai-Hai region of China, HY23×DF12（SAAS-01）, HY31×KN176（SAAS-02）and HY31×Sunoleic95R, three cross combinations were made using two elite peanut cultivars Huayu 23 and Huayu 31 as female parents, and three high oleate peanuts Sunoleic95R, DF12 and Kainong176 as male parents. One cross, four backcrosses and one selfing were carried out within three years. The progenies of each cross were examined by CAPS (cleaved amplified polymorphic sequence) and sequencing of PCR products. KASP (kompetitive allele specific PCR) was performed for more efficient examining of BC4F2 genotypes. The numbers of BC4F2 seeds with aabb genotype of three combinations SAAS-01, SAAS-02 and SAAS-03 were 1, 18 and 26, respectively. Different numbers of BC4F2 seeds with Aabb, aaBb and AaBb genotypes were obtained. These seeds could be grown and selfed to obtain the aabb homozygous. These materials could provide useful resources for breeding of high oleic acid peanut cultivars with high yield and suitable for Shandong and Huang-Huai-Hai region.
To study the relationship between endogenous hormones, photosynthetic characteristics and the cold resistance of Brassica rapa, 6 cultivars with different cold resistance were investigated on gibberellic acid (GA3), auxin (IAA), corn nucleoside hormone (ZR), abscisic acid (ABA) and photosynthetic characteristic parameters by ELISA (enzyme-linked immunosorbent assay) and CIRAS-2 at 25oC, 12oC, 8oC, 3oC, −1oC and −4oC. Results showed that contents of ABA, IAA, GA3 and ZR, and their ratios (ABA/GA3, ABA/IAA and ABA/ZR) in leaves reached highly significant level (P <0.001), which indicated temperature and cultivar had great influence on hormone contents and their balance. Cold and freezing stress reduced synthesis of IAA, GA3 and ZR in both winter and spring type rapeseed. The degree of spring type decrease was greater. When temperature came down, ABA content increased in winter type, but decreased in spring type. And the proportion of increase in strong cold resistant rapeseed was higher than that of weak cold resistant ones. Cold stress also decreased photosynthetic rate (Pn) and stomatal conductance (Gs). The decreasing speeds of winter-type and strong cold resistant cultivars were slower than those of spring type and weak cold resistant ones. The ABA content, the ratio of ABA/GA3, ABA/ZR and ABA/IAA had significant negative correlation with both Pn and Gs under low temperature. It indicated that dynamic balance between ABA and IAA, GA3 and ZR had significant effects on cold tolerance and photosynthetic efficiency of B. rapa.
To efficiently evaluate cadmium (Cd) tolerance of Brassica napus seedlings, 13 germplasm accessions were studied on morphological responses to 6 different Cd stress concentrations (0, 1, 2.5, 5, 10, 20, 30 mg/L cadmium solution). Results showed that after Cd stress, rapeseed seedlings turned dwarf, taproot and lateral root became shorter with less lateral roots, had smaller leaf area and more yellow leaf. 5mg/L Cd solution was selected as an optional concentration. 9 phenotypic traits (including relative root length, relative stems length, relative plant length, relative root green weight, relative stems green weight, relative plant green weight, relative root dry weight, relative stems dry weight, and relative plant dry weight) were selected as Cd tolerance evaluation indexes. Multiple indicators method was more effective than single index method on evaluation, because it integrated all phenotypic traits responding to Cd stress. Among the methods, principal component analysis was the best method compared to Subordinate function value method. By 3 principal components (with APC 0.92), materials were classified into Cd tolerance group and Cd sensitive group.
To reduce the error of two-dimensional projection on live, curly and deformed leave measurement, Kinect sensor was used to capture three-dimensional data with color and depth images of rape leaves. By using color image, the leaf was separated from the background according to its conspicuous gray-value in green channel. The color image without background was mapped to depth image. By using calibration square in depth image, the pixel- physical scaling parameter was calculated for 3D point cloud of the leaf. Contour of the leaf was detected and smoothed first by point cloud, and then the cloud was mapped meshing with interpolation. Finally, the total area of the leaf including grids areas were counted by Helen formula. Results show that Kinect sensor method had the advantages of low cost, high precision and universal application. The coefficient of the positive correlation between measured value and real value was 0.998 5. Regression equation y=0.955x+22.357 could be used to correct the measured value to reflect the actual value.
In order to explore the morphological response of strip cropped soybean to different light environment on two sides, soybean cultivar Nandou 12 were shaded with shade net by one side and both sides, respectively. Main stem length, stem diameter, pitch number of stems, leaf area, specific leaf area and biomass accumulation were measured at the 0th, 10th, 20th, and 30th days after shade. Results showed that elongation, thinness, and pitch number of main stems of soybean significantly decreased under unilateral shade and bilateral shade. After unilateral shade treatment, the leaf area of soybean significantly increased, so did the specific leaf area. Moreover, the specific leaf area on shaded side was larger than that on non-shaded side, but both smaller than that of bilateral shaded plants. Besides, unilateral shade treatment also caused the decline of biomass of plant, stem, leaf and petiole. And the biomass on shaded side decreased more than that on the non-shaded side, but less than that of bilateral shaded plants. In conclusion, the shade responses could be found in soybean under unilateral shade treatment, on both shaded and non-shaded side. The non-shaded side of unilateral shade treated plants displayed a compensatory effect on morphology of the shaded side, showing an overall adaption to different light environment on two sides of the plant.
To develop a rapid seed soaking technology of peanut pods, the optimum vacuum pressure and treatment time were determined. The results showed that the germination rate of the peanut increased with the extended soaking time under 0.06 Mpa vacuum pressure, while decreased under 0.08 Mpa and 0.1 Mpa vacuum pressure. With increasing of the treatment time, the electrical conductivity increased under 0.08Mpa and 0.1Mpa. The optimal treatment condition for seed soaking of peanut pods was 0.06 Mpa vacuum pressure for 15 min. The emergence rate and economic contributors of pods-sowing treated with vacuum pressure were superior to seeds-sowing in the field experiments.
To establish an efficient identification and evaluation method of rapeseed nitrogen nutrition efficiency, greenhouse pot experiment was conducted using 75 germplasm accessions under 3 nitrogen levels in soil (0.05, 0.2, 0.3 g/kg). 15 seedling traits were measured to assess variations of nitrogen nutrition efficiencies including nitrogen absorption efficiency (NAE) and nitrogen utilization efficiency (NUE). Results showed that the sensitivity of different traits to different nitrogen levels was different. Genetic variations of root surface area under different nitrogen treatments were the greatest. Under low nitrogen stress, the differences increased among the accessions. Nitrogen nutrition efficiency of rapeseed genotypes at seedling stage showed significant difference. Distribution patterns showed that both high and low efficiency materials were rare. Low nitrogen treatment (0.05 g/kg) was the best nitrogen application for identification and evaluation on NAE at seedling stage. The indirect indexes were aboveground plant dry weight and root-shoot rate. High nitrogen treatment (0.3 g/kg) was the best nitrogen application for identification and evaluation on NUE at rapeseed seedling stage. The indirect indexes were aboveground plant dry weight.
To investigate the effects of N deficiency on nutrients uptake of rapeseed (Brassica napus L.), cultivar Zhongshuang 11 was planted in hydroponics with two N treatments for 14 days. Plant phenotype, biomass, nutrients content and nutrient uptake-related gene expressions were investigated and determined. Result showed that N deficiency extremely repressed rapeseed growth, decreased shoot biomass with thin, chlorotic leaves and purple stems. Nitrogen deficiency promoted root growth with higher root/shoot ratio. The effects of N deficiency on macronutrients uptake were evaluated in different rapeseed tissues. Result showed that N concentration and content in leaf, stem and root were drastically decreased under N deficiency. Phosphorus (P) concentration in leaf and stem were increased by N deficiency, but P content was decreased in root and stem. P content in leaf was not affected by N deficiency. Potassium (K) concentration and content in different tissues were all decreased by N deficiency. In addition, N deficiency notably down-regulated the expressions of K channel genes and 3 root-specific K transporter genes. It suggested that N deficiency significantly influenced the expression of genes related to K uptake and transportation, which might result in decreased K content of rapeseed plants.
In order to excavate the supply ability of calcium (Ca) by soil tillage for peanut (Arachis hypogaea L.), four treatments of no tillage (NT), shallow plough (SP), deep plough (DP), and deep loosing (DL) were carried out in three typical brown soil experimental sites to investigate the Ca nutrition function and its synergistic absorption characteristics with phosphorus (P). Results showed that (1) Ca absorption amounts in pod and above ground parts (stem, leaf and peg) were the highest under DP and SP treatments, which were increased by 12.8% ~ 41.8% compared with NT and DL treatments. (2) Ca absorption amount in peanut was not linearly related to soil Ca content under various tillage measures. Ca absorption amount were increased with soil exchangeable Ca content when it was below the threshold value of 2.7 g/kg. (3) There was significant positive correlation between the absorption of Ca (Ca2+) and P (H2PO4–/HPO42–). The synergistic absorption effect of Ca and P was better in pod than in parts above ground. (4) Peanut pod yield was increased by 610 kg/hm2 and crude protein was increased by 108 kg/hm2 when Ca absorption amount in pod increased by 10 kg/hm2. In general, maintaining the appropriate content of soil exchangeable Ca by plough measures with 20~30 cm contributed to the synergistic absorptions of Ca and P, which would significantly improve the yield and quality of peanut.
A susceptible cultivar Liaodou 15 and a resistant cultivar Huipizhi heidou (ZDD2315) were selected to research the gene expression of calmodulins from soybeans roots after SCN3 infection. RNA from infected and uninfected roots was extracted at 1d, 3d, 5d, 9d, 15d and 20d and real-time PCR was carried out respectively. The complete SCaM4 gene was amplified into binary expression vector to construct Pcombia1303- SCaM4. LSCM was used to observe the location. The results demonstrated that in both cultivars the five calmodulins genes expression levels reached the maximum at the fifth day after infection among six time points. The gene expression of SCaM4 after infected was 1.85 fold of uninfected. And in resistant cultivar the relative expression of SCaM4 was 1.96 times of that in susceptible cultivar. The result of subcellular localization showed that SCaM4 protein was located on the cytomembrane. In summary SCaM4 may participate in the interaction of soybean and SCN3.
Peanut scab caused by Sphaceloma arachidis is one of the important leaf diseases of peanut, distributed widely in major peanut production areas in China. In order to explore the mechanism and role of host defensive enzymes in plant-pathogen interaction, the infectious condition of S. arachidis and the dynamic changes of defense enzymes activity in different resistance peanut varieties were investigated. The results showed the incubation period of peanut scab was 120h in susceptible variety Baisha 1016 at 25°C-30°C, was 240h at 15°C. Infection ability of fungi had significant positive correlation with time in moist chamber. The shortest time in moist chamber was about 4h. The optimum condition for infection was 25-30°C and 42-48h in moist chamber respectively. Infection ability of S. arachidis had negative correlation with their culture time. Activities of defense enzymes including phenylalanine ammonia-lyase, peroxidase, polyphenol oxidase, superoxide dismutase and catalase in two peanut varieties all increased after inoculation and then decreased. Enzymes activities in resistant varieties (Fuhua 17) peaked at 24 to 36h post inoculation while in Baisha 1016 was at 48 to 72h post inoculation. The disease severity and infection index of Baisha 1016 were significantly higher than that of Fuhua 17 infected with S. arachidis.
In order to clarify weed diversity and characteristics of weed communities in peanut fields of main growing regions in Hubei province, field weed survey was conducted using an inverted W-pattern with 9 sampling points. A total of 76 weed species belonging to 24 families were found, including 4 dominant species, 6 regional dominant species, 10 minor species, and 56 sporadic species. The relative abundance (RA) of Digitaria sanguinalis, Eclipta prostrate, Commelina bengalensis and Acalypha australis were higher than those of other weed species. They were the dominant weeds in all main peanut regions in Hubei province. The most common weed communities were Eclipta prostrate + Digitaria sanguinalis + Commelina bengalensis in peanut fields in northeastern Hubei province, and Digitaria sanguinalis + Acalypha australis + Commelina bengalensis + Eclipta prostrata in northern Hubei province. The variety and diversity of weeds in peanut fields in northeastern Hubei were greater than those in northern Hubei, but the Simpson index was relatively lower.
To reduce the cost of single-chain antibody for aflatoxin and increase the expression activity of single-chain antibody, the recombinant plasmid containing the aflatoxin scFv gene pCANTAB 5E-scFv1A7 was cloned to obtain scFv gene fragment and construct a yeast expression vector pPICZαA -scFv1A7. Then SacⅠ enzyme was linearized into Pichia pastoris X-33 chromosome genome to construct pPICZαA-scFv1A7 X-33 recombinant yeast. Single-chain antibody was successfully expressed with 0.8% methanol induced secretion. Indirect competitive ELISA method was used to detect aflatoxin B1 and revealed the detection inhibition rate (IC50) of 4.5 ng/ mL, which indicated that the recombinant expression product scFv had good antigen binding activity and could be applied in diagnosis of aflatoxin contamination.
Using defatted pine nut as raw material, the effects of the extraction methods on properties of protein were compared. Pine nut proteins extracted from the defatted pine nut powder by Osboren and alkali method were comparatively evaluated for their physical-chemical characteristics and functional properties. Comparisons and analysis on Isoelectric point, thermo-stability, amino acid composition, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, SDS-PAGE, protein solubility, foaming capacity and stability, emulsifying activity and emulsion stability of different proteins were conducted. The results showed that the contents of albumin, globulin, gluten by Osboren were 77.87%、68.23%、79.05% respectively; the content of protein prepared by alkali extracted was 83.50%; Isoelectric point were 3.4、3.8、3.6、4.0. Thermo denaturation temperatures were 94 ℃、81 ℃、98 ℃、112 ℃ respectively; the essential amino acid composition of these 4 proteins were reasonable, and the Glu and Arg was the main contributing amino acid. The bands of these 4 proteins had subtle differences by SDS-PAGE, which mainly concentrated in the range of 9.31 to 41.32 ku. Functional properties of these 4 prepared proteins, excluding protein solubility, foaming capacity and stability, albumin was the best; protein’s emulsifying activity and emulsion stability by alkali method was better than others. Therefore different methods could result in differences in physical-chemical characteristics and functional properties of proteins.
To compare amino acid difference between black and white sesames, 40 samples of each sesames were collected from the main producing areas in China. Under optimized experimental conditions, amino acid compositions of both sesame samples were analyzed by hydrochloric acid hydrolysis and automatic amino acid analyzer. Results indicated that both samples contained 17 essential amino acids. Arginine and glutamic acids were dominant in both seseames. In black seseame, arginine and glutamic acids contents were 2.82%±0.26% and 4.82%±0.51%. In white seseame, their contents were 2.86%±0.24% and 4.81%±0.53%. By t test, contents of methionine and lysine in both samples were significantly different (P<0.05).
In this study, we examined the changes of fucoxanthin content and transcript quantity of key genes encoding biosynthesis pathway of fucoxanthin containing six key enzymes，phytoene synthase, phytoene desaturase, ζ-carotene desaturase, carotenoid isomerase, lycopene β-cyclase, and zeaxanthin epoxidase (excluding the unidentified enzymes in diadinoxanthin cycle and violaxanthin cycle) in the diatom P. tricornutum. P. tricornutum cells were exposed to high light intensity of 500μE/(m2•s) for 3h, 6h and 12h; and fucoxanthin content was quantified by HPLC. Fucoxanthin content increased and then declined during 12 h of high irradiance, and peaked at 6h (1.85 mg·g-1DCW), which was 2.16-fold higher than that of control sample (0h for high irradiance). Quantitive RT-PCR showed that the transcript abundances of genes encoding zeaxanthin epoxidase (zep1) and phytoene synthase (pys) peaked at 6h of high irradiance, which was two-fold higher than that of control (0h for high irradiance) and consistent with the trend of change of fucoxanthin content. The transcript quantity of the genes encoding ζ-carotene desaturase3, (crtiso3), phytoene desaturase (pds) and ζ-carotene desaturase (zds) decreased greatly upon high irradiance for 3h compared to the control, and showed a rapid response to high irradiance stress. The expression levels of genes encoding lycopene β-cyclase (lcyb) and carotenoid isomerase (crtiso1) was only enhanced after high irradiance for 12 h. The response of the expression of lcyb and crtiso1 to high irradiance stress was at least 6 h later than that of zep1 and pys. This study revealed the change of fucoxanthin content and expression pattern of genes encoding fucoxanthin biosynthesis pathway under the condition of high irradiance stress in the diatom P. tricornutum, showing an evident linear relationship between fucoxanthin content and the expression levels of zep1 and pys.
In order to obtain functional genes in response to drought stress, a DGE (digital gene expression) profile, two drought-tolerance and sensitivity materials under drought stress, had been constructed in our previous study. A gene encoding NAC transcription factor, named as GmNAC8, was chosen as candidate gene from DGE profile. The gene, the CDS full-length sequence (1 092bp), encodes 363 amino acid. By software analysis, the protein with a molecular mass of 41.82kD and PI 8.51, the N-terminal aa residue of GmNAC8 contains a typical NAC conserved domain with 42 aa, and C-terminal aa residue is highly variable. The homology tree demonstrated that GmNAC8 shared high homology with Phaseolus vulgaris, Vigna angularis and Vigna radiata. Subcellular localization revealed that GmNAC8 was expressed in the nucleus. Transcription analysis by qRT-PCR showed that the expression of GmNAC8 in leaf was apparently higher than that in root from transgenic lines, and GmNAC8 was up-regulated by 0.1mmol/L IAA, and ABA was down-regulated by 0.1mmol/L GA and SA. The leaves of transgenic Arabidopsis thaliana lines were greener than CK, and drought tolerance was improved under water-deficient condition by overexpression of GmNAC8. The drought-tolerant physiological analysis revealed that the contents of protein, proline, and POD activity were higher in transgenic lines than CK in 10 day drought treatment, while malonaldehyde (MDA) was significantly lower than CK, only occupied 42%. This study would provide better understanding of the molecular mechanisms of plants response to drought.
