• 2020 Volume 42 Issue 1
    Published: 28 February 2020

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  • CHEN Qing-shan, JIANG Hong-wei, XIN Da-wei, ZHU Rong-sheng, LIU Chun-yan, LI Can-dong
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     Backcross introgression lines are genetic and breeding materials constructed by backcrossing andmarker-assisted selection. After multiple backcrossing, the progenies only contain one or a few chromosome seg‐ments of the donor parents in the genetic background of the recurrent parents, so they can be used as important ma‐terials for QTL analysis. At the same time, multi-generation backcross is beneficial to break the linkage between ex‐cellent gene and bad gene, and good gene is introduced into the recurrent parent material with good performance,and further improve breeding material. In view of their consistent genetic background, introgression lines play an im‐portant role in fine mapping and gene cloning, QTL interaction, genetic validation, crop pyramiding breeding andbreeding by molecular design. In this paper,the advanced progresses on the construction and application of CSILpopulation in crop were reviewed.

  • JIANG Hong-wei, LI Can-dong , LI Rui-chao , LI Ying-ying , YIN Yan-bin , MA Zhan-zhou , ZENG Qing-li , ZHANG Wen-bo , LIU Chun-yan , CHEN Qing-shan
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     To excavate the excellent rare gene of wild soybean, this study had been carried out since 2006, Sui⁃ nong 14 (recurrent parent) and wild soybean ZYD00006 (donor parent) were used as parents to construct a set of chromosome fragment introduction lines (substitution lines) covering the whole genome of wild soybean through hy⁃ bridization, backcross and marker assisted selection. A total of 192 plant rows in this population, included 237 wild soybean introduction fragments, with the average number of introduction fragments per linkage group of 11.85. The total length of the introduction fragment was 1865.17cM with the coverage of 83.23% of wild soybean genome. The coverage rate of linkage group L was the highest of 100%; linkage group N had the least coverage, of 53.17%. The maximum length of segment was 43.30 cM and the minimum one was 0.22 cM. Highly consistent genetic back⁃ ground was of great significance for excavating important genes of soybean and wild soybean-specific excellent gene. At the same time, the introduction of wild resources greatly enriched the genetic basis of cultivated soybean, and further enriched the phenotypic variation of the introgression lines, providing an important material basis for soybean genetic breeding. 
  • TIAN Bo-yu, SUN Zhi-jun, LIU Han-xi, WEN Ying-nan, WANG Jie-qi, LYU Hao, ZHOU Hong-yang, HUANG Xiu-zheng, LIU Chun-yan, WANG Jin-hui, XIN Da-wei , CHEN Qing-shan
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    In this study, TtsI, a regulator of HH103 type III secretion system in Sinorhizobium was detected. The mutant HH103ΩTtsI was obtained by three-parental hybridization and identified by PCR. Type III effector ex⁃ pression analysis showed that TtsI mutation could inhibit the transcription and expression of effector genes such as NopAA, NopD, NopL, and NopP in the presence of genistein. When TtsI mutated, it could significantly inhibit the synthesis of rhizobium HH103 type III effector. TtsI mutants and wild-type rhizobium HH103 were used to identify nodules in 10 soybean germplasm of different ecological regions. These results showed that the TtsI mutation could significantly reduce the average number of nodule and the dry weight of nodule. The nodule phenotype identification results of the introduced line population showed that the TtsI mutant significantly reduced the average nodule num⁃ ber of the introduced line. This showed that TtsI played an important role in the establishment of a soybean-rhizobia symbiotic system depending on other type III effectors.
  • SUN Zhi-jun, CAI Ying-bo, LYU Hao, WANG Jie-qi, LIU Han-xi, WEN Ying-nan, LIU Chun-yan, WANG Jin-hui, XIN Da-wei , CHEN Qing-shan (
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     Nitrogen is an important nutrient for plant growth, and biological nitrogen fixation is an important source of nitrogen to soybean growth. In this study, we used Suinong14 as the recurrent parent and wild bean ZYD00006 as the male parent to construct an introgression lines covering the whole genome of wild soybean. QTL mapping of nodule number and nodule dry weight based on soybean introduced line group. Three QTLs for nodule dry weight, located on three linkage groups of N, M and D2. Five QTLs for nodule number, located on linkage groups K, F, J and D2. There are overlapping segments (7.20-7.79Mb) on D2 linkage group. Gene annotation was performed on 63 genes in this overlapping segment, and 6 genes related to symbiosis and disease resistance were se‐ lected as candidate genes for further verification. The qRT-PCR analysis showed that the expression pattern of Gly⁃ ma.17G097000 during rhizobial infection was very different compared to the control. Glyma.17G097000 belongs to the GmHIR gene family, and 11 family members were found in the soybean genome. The structural similarity of Gm⁃ HIR family genes is very high, and genes expression is tissue-specific. Soybean HIR protein has two domains, Stomatins and Prohibitin, and can participate in physiological processes such as ion channel regulation, which are related to plant disease resistance and cell cycle. The GmHIR genes are derived from four ancestors and highly con‐ served during evolution. The qRT-PCR analysis of 11 members of GmHIR gene family showed that the expression patterns of these three genes Glyma.05G029800, Glyma.09G154400 and Glyma.17G097000 during rhizobial infec‐ tion were significantly different from the control. These results indicated that these three genes could have played an important role in the establishment of a soybean symbiosis system, and participated in the regulation of ion channels and immune responses during the establishment of a symbiotic system of rhizobium and soybean. This study lays the foundation for the study of soybean-rhizobia symbiosis mechanism and provides effective candidate genes.
  • LI Jian-yi, WANG Yue, XIN Da-wei, LI Rui-chao, YU Jing-yao, HUANG Shi-yu, WANG Xin-yu, WU Xiao-xia , CHEN Qing-shan
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    Nitrogen is one of the many elements necessary for plant growth. Symbiosis of soybean rhizobium can provide sufficient nitrogen for soybean. It is important to study the mechanism of soybean rhizobium symbiosis and excavate candidate genes for controlling symbiosis. Given that the QTL for regulating soybean symbiosis is large at this stage, it cannot be directly applied to practice. In this experiment, 68 QTLs for soybean nodulation were col⁃ lated, and they were optimized by Overview and colinearity analysis. A high-confidence QTLs interval was obtained on the chromosome Gm06. The gene was annotated and 43 genes were obtained, including one coding system C2 Calcium / lipid binding and GRAM domain-containing proteins, a side root-forming protein and an E3 ubiquitin li⁃ gase-related protein, and it’s a so possible to search for strains with inserts at this location in the CSSLs population for identification of nodulation traits. The results showed that 15-15.5 Mb on the C2 linkage group was crucial for the symbiotic nodulation of soybean rhizobium.
  • LI Can-dong, JIANG Hong-wei , GUO Tai , WANG Zhi-xin , ZHENG Wei , ZHANG Zhen-yu , ZHAO Haihong , WANG Nan-nan , CHEN Qing-shan
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    In order to identify stable and universal QTLs for the number of four-seed-pod trait on soybean in different years and groups, based on the phenotypic data in 2013-2016 year, a RILs population and its SLAF-seq high density genetic map were used to identify the QTLs. Phenotypic validation had been done with introgression lines carrying the target interval. The results showed that there were 8 QTLs in two chromosome identified. Four QTLs in Gm06 chromosome had close position and positive additive effect. The target interval size was 0.62Mb. Four QTLs in Gm16 chromosome had same negative and positive additive effect. The target interval size was 1.04Mb. These QTLs were stable in different years. 5 introgression lines carrying the target interval of Gm06 chro⁃ mosome had significantly higher phenotypic value than recurrent parent in different years. 7 introgression lines car⁃ rying the target interval of Gm16 chromosome had significantly lower phenotypic value than recurrent parent in dif⁃ ferent years. The results showed that the target interval introgression led to synergistic or deleterious effects of the phenotypic of the number of four-seed-pod trait. The accuracy and commonality of QTLs were verified in genomewide introgression lines. The results of this research provided theoretical foundation for candidate gene mining and molecular assisted breeding of the number of four-seed-pod trait on soybean. 
  • ZHAO Yuan-yuan , LI Rui-chao, JIANG Hong-wei , WANG Qiao , XIE Jian-guo , LIU Chun-yan , WU Xiao-xia , CHEN Qing-shan
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    The first pod height is a key indicator of soybean varieties suitable for mechanical harvest. Varieties with lower first pod height may cause part of the plant to be cut or missed during mechanical harvesting, resulting in total yield loss. Therefore, the candidate genes related to the first pod high traits in soybean is very important for soy⁃ bean mechanization breeding. In this study, the Inclusive Composite Interval Mapping (ICIM) method was used to map the QTLs of soybean first pod height traits for 208 chromosome segment substitution lines (CSSL), and obtained 9 QTLs related to soybean first pods height which were distributed on 8 linkage groups. Combined with the BSA re⁃ sequencing results, QTLs related to soybean first pod height were mapped to 1.1Mb on the C1 linkage group and 0.05Mb on the L linkage group, and gene annotation was performed. Through gene annotation database and informat⁃ ics analysis, five candidate genes related to soybean first pod height were obtained in two consensus QTL intervals. These results provided theoretical basis for fine QTL mapping of soybean first pod height and selective breeding of mechanized high-quality and high-yield soybean varieties. 
  • ZHU Rong-sheng , LI Shuai , YAN Xue-hui , YAN Zhuang-zhuang , YU Jiang-lin , SHI Jia , SU Xiao-yan, CAO Yang-yang , XIN Da-wei , LI Yang , JIANG Hong-wei , ZHAO Zhen-qing , ZHANG Zhan-guo , CHEN Qing-shan
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     The construction of soybean imported line population and selection of materials are important in soy⁃ bean breeding. The selection of imported line materials for a specific phenotype can not only improve the efficiency of breeding, but also speed up the process of locating target trait genes. In this study, the semantic segmentation technique in machine detection was used to automatically extract the main stem-related phenotypes from the import⁃ ed lines (190 individuals) constructed for both parents, Suinong 14 (recurrent parent) and wild soybean ZYD00006 (donor parent). On this basis, this study systematically clustered the tested materials for the main stem-related phe⁃ notypes, and the results clearly showed the aggregation of the tested materials (near-equal materials) based on the main stem-phenotypes. It became an important basis for the selection of materials for the main stem-phenotypes. At the same time, the results could greatly accelerate the locating procedure of the main stem related phenotype genes. 
  • LIU Ren-feng , HUANG Shi-yao , NIE Yong-peng , XU Sheng-yong
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    Silique observation and measurement are essential for rapeseed breeding. In this paper, an automat⁃ ic detection method was proposed to replace the traditional manual method. A device was designed to acquire video of scattered siliques, based on pulling and vibrating stacked siliques. Silique videos were extracted to frames, and then, by using QR code as marker blocks in image, the key frames containing all siliques were effectively extracted and spliced into individual intact. Crossed siliques in frame caused an error in measurement. A cutting method of crossed siliques image based on concave point extraction and matching was proposed. By this method, all kinds of crossed siliques could be identified with accuracy rate of 98.0%. In the measurement of phenotypic parameters, a core diameter Otsu method was proposed to judge the posture of silique, by which elliptic long and short axis of the cross section of the silique was estimated, and then length, surface area and volume of the silique were calculated. Results demonstrated good accuracy and adaptability to different varieties of rape by this method. Estimation error of length, surface area and volume were less than 2.9%, 4.8% and 5.0% respectively. Thus the method could be an effective replacement of artificial way and provide key basic data for rapeseed and agricultural research fields.
  • YU Er-ru, LI Hui-lin, YANG Hang, YANG Sheng-xian, FENG Bin, WEI Zhong-fen
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     For better evaluate and utilize perilla resources in Guizhou, 274 accessions were collected and clas⁃ sified to find out their features and differences. Among them, main seed quality traits of 188 resources were investi⁃ gated. Results showed that perilla resources were widely distributed throughout the province, and most of them were collected in the south, southwest and Bijie City of Guizhou, which together accounted for 55.11% of the total re⁃ sources. The average oil content and protein content of the tested perilla resources were 36.69% and 25.96% respec⁃ tively. The main component of the fatty acids was linolenic acid, with an average of 59.27%. Correlation analysis of the 7 main quality traits showed a significant positive correlation between oil and protein (r= 0.43, p< 0.01) and a significant negative correlation between linolenic acid and the other 4 fatty acid components (r= -0.70 to -0.52, p< 0.01). By cluster analysis, the tested resources were classified into V types, regarded as OH PH LH (high oil, high pro⁃ tein, high linolenic acid) -type, OH PH LL (high oil, high protein, low linolenic acid)-type, OL PL (low oil and low pro⁃ tein)-type, medium quality-type and OUHLUH (ultra high oil-ultra high linolenic acid)-type respectively. The study clarified the regional distribution and quality characteristics of Guizhou’s perilla resources, and could benefit the researchers in high efficiency utilization and quality breeding of perilla. 
  • DONG Cheng, YANG Jing-song, LIU Zi-yan, LI Hai-long, CHU Zhuo, WEN Qiu-ju, XIE Zi-yang, LIU Shu-ming, GUAN Li-li
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     Dehydrates was a member of the group 2 late embryogenesis abundant protein family, which were
    high disorder proteins and responded to adversity stress. Based on the transcriptome data of safflower under drought
    stress, CtDHN1, a dehydrin gene respondimg to drought stress was identified and cloned by using RT-PCR tech⁃
    niques, which encoded 318 amino acids with the molecular weight of 31.1 kDa and the pI value of 6.36. Bioinformat⁃
    ics analysis indicated that CtDHN1 protein belonged to YnSKn-type dehydrin. Prokaryotic expression analysis re⁃
    vealed that CtDHN1 protein was a soluble protein. The protein had the highest expression level when the concentra⁃
    tion of IPTG was 0.4 mmol/L, the OD600 value was 0.8, the induction time was 4 h, and the induction temperature
    was 37℃. The E. coli stress tolerance analysis demonstrated that CtDHN1 gene tolerated a drought and a high salt
    environment of 1.5 mol/L sorbitol and 1.3 mol/L sodium chloride. In vitro enzyme activity assays indicated that Ct⁃
    DHN1 protein protected LDH enzyme activity in 500 mmol/L and sorbitol 300 mmol/L sodium chloride. This re⁃
    search provided a theoretical basis for the molecular mechanism of CtDHN1 gene in safflower resistance to abiotic
  • XU Yao-zhao, SUN Wan-cang , FANG Yan, SUN Bo-lin
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     To clarify the relationship between material transport, photosynthesis, fluorescence dynamics and cold tolerance of winter turnip rape (Brassica rapa L.)and to further provide theoretical basis for rape breeding and evaluation of cold resistance, 2 cultivars Longyou 7 (strong freezing-tolerance) and Tianyou 4 (weak freezing-toler⁃ ance) were determined in natural temperature falling. Effects of low temperature on dry matter distribution, photo⁃ synthetic performance and kinetic curve of chlorophyll fluorescence induction were investigated under field condi⁃ tions. It was shown that dry matter and distribution rates of dry matter in roots of both cultivars increased, but photo⁃ synthetic rate, transpiration rate, Fo (fluorescence at O-step at 20 microseconds), Fk (fluorescence at K-step, 300 microseconds), Fj (fluorescence at J-step at 2 millisecond) and Fm (dark-adapted maximum fluorescence at P-step) of chlorophyll fluorescence induced kinetics curves of O-J-I-P decreased, with the decreasing temperature. Thus the decline of photosynthetic performance might help accumulation of dry matter in roots at low temperature. Photo⁃ system damage and photo inhibition occurred, which presented with falling of PSII receptors (area), photosynthetic performance index (PI), maximum fluorescence (Fm) and maximum photochemical efficiency (Fv/Fm), except the ris⁃ ing of initial fluorescence (Fo). Longyou 7 had lower Pn, Fm, Fv/Fm, and had lower potential activity of PSⅡ (Fv/ Fo) and reduced capacity of QA and QB. It showed weaker photosynthetic capacity and stronger photo inhibition in Longyou 7 than those of Tianyou 4 under low temperatures. These indicated that the photosynthetic capacity of Longyou 7 decreased at low temperature before winter, which might be due to non-stomatal factors, and low temper⁃ ature might lead to enhanced photoinhibition. In summary mary, the cold resistance of winter turnip rape was relat⁃ ed to leaf photosynthetic characteristics and dry matter accumulation under low temperature. Weak photosynthesis could reduce growth activity of aboveground parts and root matter consumption, which was conducive to dry matter accumulation of rape root to enhance its cold resistance.
  • MA Jia-bin , YU Xiao-bo , WU Hai-ying, ZHANG Ming-rong
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    To investigate the soybean nitrogen fixation ability response to the inoculation of different rhizobi⁃ um, two soybean varieties, Nandou 20 ( ND20 ) and Nanxiadou 25 (ND25), were inoculated with Bradyrhizobium ja⁃ ponicum 5038 and B. japonicum 5136 in this field experiment. The photosynthetic traits, chlorophyll content, nodule number and the physiological index of fresh nodule, such as sucrose content, leghaemoglobin content and nitroge⁃ nase reductive ability were investigated. The result revealed that chlorophyll content (chl), photosynthetic rate (Pn) and stomatal conductance (Gs) increased significantly but chla/b value decreased significantly after inoculation. Nodule number, nodule biomass, shoot and root biomass all increased significantly, biomass per nodule and root/ shoot value changed without significace. Sucrose content, leghemoglobin content and nitrogenase reductive ability of nodule all increased significantly. Compared with ND20, ND25 had bigger increasing amplitude, not only the photo⁃ synthetic characteristics but also the nodule characteristics and nitrogenase reductive ability.
  • LI Shu-yu , XIONG Jie , DING ge , CHEN Lun-lin , ZOU Xiao-yun, ZOU Xiao-fen , SONG Lai-qiang
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    Under rice-rapeseed rotation in the Yangzi River Basin, risk of rice herbicide residues on rapeseed production should be evaluate for selection of herbicide-tolerant rapeseed. Thus 19 rapeseed varieties were evaluat⁃ ed under pyrazosulfuron, pretilachlor, bentazone and cyhalofop herbicides stresses. Rapeseed traits were investigat⁃ ed on seed germination rate, plant height, maximum root length, fresh weight aboveground, fresh weight of roots, dry weight aboveground and root dry weight. Results showed that, under herbicide stresses, the tolerance coefficients of different genotype were significantly different. By cluster analysis, varieties were selected with relatively strong tol⁃ erance to different herbicides. It showed that comprehensive evaluation of herbicide tolerance was effective on rape⁃ seed by integration of principal component analysis, subordinative function value and clustering analysis, which could avoid bias and instability of single index analysis. It suggested a practical selecting method for rapeseed as succession crop after rice according to actual use of herbicides in rice plantation. 
  • YU Jing , WU Ju-xiang , XU Man-lin , ZHANG Xia , GUO Zhi-qing , CHEN Dian-xu , ZHANG Zhi-meng , LI Xin-guo , CHI Yu-cheng , WAN Shu-bo
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     Peanut pod rot is one of the most important soil-borne diseases on peanut. It will cause pod rot, and the seriously affected field can lead to total crop failure. In recent years, Pythium myriotylum was identified as the main pathogen of peanut pod rot in the major peanut producing areas of China. In order to control this disease effi⁃ ciently, 6 fungicides were screened indoors, the experimental results showed that 96.8% fludioxonil had best inhibi⁃ tory effect on mycelial growth of P. myriotylum. The EC50 value was 3.16 × 10-5 mg/L. The six fungicides were used for root irrigation, after potted seedlings inoculation, and the disease incidence was investigated after the pods rip⁃ ened. The results showed that 96.8% fludioxonil was the best potential chemical agent for preventing peanut pod rot.
  • LI Wen-bo, GAO Yu, CUI Juan, SHI Shu-sen
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    To ascertain the influence of temperature on development, fecundity and population of Atractomor⁃ pha sinensis (I.Bolivar) in field, duration of different developmental stages of the insect were compared in laboratory under constant temperature of 16℃, 20℃, 24℃, 28℃ and 32℃, with photoperiod of 16L:8D at RH 70%. With soy⁃ bean leaves as food, duration, developmental rate of each life stage and reproduction were systematically observed. Developmental threshold temperature and effective accumulated temperature were further calculated. Population trends under different experimental temperatures were also analyzed by life table technology. Results showed that developmental period of each stage of A. sinensis were decreased from 16 to 32 o C, and a significant positive correla⁃ tion between developmental rate and temperature was confirmed by fitting equation. Developmental threshold tem⁃ peratures of egg, nymph and adult were 9.58, 10.55 and 11.40 o C respectively. Effective accumulated temperature were 449.80, 540.03 and 430.94 degree-days respectively. Insect fecundity was the highest at 28℃, with average 66.12 eggs laid per female, followed by 60.30 eggs laid per female at 24 o C and the lowest was 28 eggs at 16 o C. Ex⁃ perimental population trend index was less than 1 under each test temperature, and the maximum was at 24 o C, which was 18.97. These results were expected to provide a scientific basis for field prediction and integrated man⁃ agement of A. sinensis (I.Bolivar). 
  • TANG Jian-rong, LIU Yue-lian , WU Ling-ping, LU Jian-nong, YIN Xue-gui
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     To clarify the cause of castor bean bud rot, pathogenic strains were isolated by tissue isolation and tested for pathogenicity. Classification status of the pathogens were determined by ITS-PCR combined with morpho⁃ logical identification. Results showed that RFR1-4 isolated from the disease samples, was confirmed to be the pathogen according to Koch's rule. It was identified as Fusarium solani by morphological characteristics. By ITS analysis, EF1-α and β-tubulin sequences revealed a 100% identity with F. solani species complex by polyphasic identification of FUSARIUM-ID database. Therefor, the pathogenic strain (RFR1-4) was identified to be a member of F. solani species complex which caused castor bean bud rot. Key words: castor;bean bud rot;Fusarium solani species complex 蓖麻(Ricinus communis L.
  • ​WANG Zhao-zhao, , LU Feng-yin , ZHU Guang-cheng , YANG Hui , GUO Wen-ying , ZHAI Chen-lu , HAN Jun-hao , DONG Jun-hui , DONG Tie-you
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     In order to improve the quality of dried peanut pods, we had coupled hot air with pulse intermittent low-intensity microwave, high-intensity microwave, and high & low intensity microwave respectively in the drying process. Drying characteristics, peanut pods quality and drying energy consumption of different drying processes were analyzed. The result showed that the best process was to combine hot air with pulse intermittent and high & low intensity microwave. With an irradiation of 0.95 W/g microwave for 16.7 min and 0.48 W/g microwave for 11.7 min, the moisture content of peanut pods was reduced from 50.0% to 15.5% in 198.3 min, and the total energy con⁃ sumption of the process was respectively 37.1% and 8.6% lower, the quality of dried peanut pods was better. There⁃ fore, analysis of the drying process which couples hot air with pulse intermittent and high & low intensity microwave is of vital importance to wastage reduction of peanut pods.
  • WANG Lei, TAN Mei-lian , FU Chun-ling, WANG Wei, WANG Li-jun, YIN Zi-yan, YAN Xing-chu
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     For rapid and high throughput prediction of the quality of sunflower seeds, calibration equations were established on 154 materials of sunflower husked seeds. Combination methods included near-infrared spec⁃ troscopy (NIRS) scanning and chemical detecting with chemometrics method to determine the optimal calibration models. Results showed that the model of modified partial least-squares method was appropriate. The determination coefficient of NIRS model for crude fat, crude protein, oleic acid, linoleic acid, saturated fatty acid and unsaturated fatty acid were 0.975, 0.950, 0.973, 0.951 and 0.913 respectively. Their cross-validation correlation coefficients were 0.969, 0.939, 0.915, 0.927 and 0.711 correspondingly. Validation testing results showed that the external vali⁃ dation correlation coefficient for crude fat, crude protein, oleic acid, linoleic acid, saturated fatty acid and unsaturat⁃ ed fatty acid were 0.959, 0.950, 0.937, 0.906 and 0.930 respectively. The results indicated that the established NIRS model could be used as a tool for rapid prediction of qualitative characteristics in sunflower husked seed for large-scale screening of sunflower quality breeding.
  • GUO Ting-ting , WAN Chu-yun , HUANG Feng-hong , CHEN Si-han
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     Subcritical fluid extraction (SFE), an efficient extraction technology, has attracted much attention in oil processing industry. In this paper, the literatures of SFE on oil extraction were briefly introduced, and the re⁃ search progress of SFE was reviewed in oil extraction from different sources and minor compounds. Furthermore, prospect of SFE was forecasted and discussed.