In order to provide basic design parameters for the industrial pyrolysis process,the transformation behavior of nitrogen was investigated using wheat straw as raw material.The distributions of nitrogen in pyrolysis ch...In order to provide basic design parameters for the industrial pyrolysis process,the transformation behavior of nitrogen was investigated using wheat straw as raw material.The distributions of nitrogen in pyrolysis char,oil,and gas were obtained and the nitrogenous components in the products were analyzed systematically by X-ray photoelectron spectroscopy(XPS),pyrolysis-gas chromatography/mass spectrometry(Py-GC/MS)and thermogravimetric-Fourier transform infrared spectrometry(TG-FTIR).The nitrogen distribution ranges of the pyrolysis char,oil,and gas were 37.34%–54.82%,32.87%–40.94%and 10.20%–28.83%,respectively.More nitrogen was retained in char at lower pyrolysis temperature and the nitrogen distribution of oil was from rise to decline with increasing temperature.The most abundant N-containing compounds in three-phase products were pyrrole-N,amines,and HCN,respectively.In addition,the transformation mechanism of nitrogen from wheat straw to pyrolysis products was concluded.展开更多
Wheat is a critical crop,extensively consumed worldwide,and its production enhancement is essential to meet escalating demand.The presence of diseases like stem rust,leaf rust,yellow rust,and tan spot significantly di...Wheat is a critical crop,extensively consumed worldwide,and its production enhancement is essential to meet escalating demand.The presence of diseases like stem rust,leaf rust,yellow rust,and tan spot significantly diminishes wheat yield,making the early and precise identification of these diseases vital for effective disease management.With advancements in deep learning algorithms,researchers have proposed many methods for the automated detection of disease pathogens;however,accurately detectingmultiple disease pathogens simultaneously remains a challenge.This challenge arises due to the scarcity of RGB images for multiple diseases,class imbalance in existing public datasets,and the difficulty in extracting features that discriminate between multiple classes of disease pathogens.In this research,a novel method is proposed based on Transfer Generative Adversarial Networks for augmenting existing data,thereby overcoming the problems of class imbalance and data scarcity.This study proposes a customized architecture of Vision Transformers(ViT),where the feature vector is obtained by concatenating features extracted from the custom ViT and Graph Neural Networks.This paper also proposes a Model AgnosticMeta Learning(MAML)based ensemble classifier for accurate classification.The proposedmodel,validated on public datasets for wheat disease pathogen classification,achieved a test accuracy of 99.20%and an F1-score of 97.95%.Compared with existing state-of-the-art methods,this proposed model outperforms in terms of accuracy,F1-score,and the number of disease pathogens detection.In future,more diseases can be included for detection along with some other modalities like pests and weed.展开更多
[Objective] The aim was to provide theoretical basis for field moisture conserving irrigation.[Method] With Xiaoyan No.6 as tested material,three different kinds of mulching irrigation treatments were carried out (st...[Objective] The aim was to provide theoretical basis for field moisture conserving irrigation.[Method] With Xiaoyan No.6 as tested material,three different kinds of mulching irrigation treatments were carried out (straw mulching;plastic mulching;PAM control adjustment mulching).With non-mulching treatment as control,moisture conserving effect of different treatments were compared.[Result] The results showed that the water consumption of winter wheat under different soil moisture conservation treatments was low at earlier stage and later stage,but high at mid-stage,which was consistent with the water consumption law of control.There were some differences in terms of consumption intensity because of irrigation schedule and growth condition;soil moisture conservation treatments could restrain ineffective evaporation of soil moisture before anthesis.We also found that the variation of soil moisture at depth of 0-20 cm in PAM and control treatment was dramatic.The soil moisture of the former was lower than the latter at the depth of 0-20 cm,but higher at the depth of 20-50 cm.The difference of soil moisture at the depth of 0-50 cm was significant.[Conclusion] Plastic mulching and straw mulching could restrain evaporation effectively.展开更多
Wheat transformation efficiency is closely related to several factors such as receptor genotype, constructed plasmid and selection procedure after bombardment or co-cultivation. In our study, several kinds of antibiot...Wheat transformation efficiency is closely related to several factors such as receptor genotype, constructed plasmid and selection procedure after bombardment or co-cultivation. In our study, several kinds of antibiotics, which were normally used in plant transformation to the selection genes of nptII, bar and hpt, were tested for the optimal concentrations for wheat transformation. The results showed that 25 - 50mg/L of geneticin (G418) was suitable for the selection of nptll, kanamycin or neomycin was not suitable for use. 3 -5mg/L of phosphinothricin (PPT) or biolaphos could be used for the selection of bar, 100 - 150mg/L of hygromycin for the selection of hpt. Yangmai 158 and Yangmai 10 with high tissue culture response and good agronomic characteristics were screened from 25 potential Chinese wheat cultivars. The concentration changing of selectable agent in selection medium was helpful to obtain enough regeneration plantlets with strong root system.展开更多
More than 20 years have passed since the first report on successful genetic transformation of wheat. With the establishment and improvement of transformation platform, great progresses have been made on wheat genetic ...More than 20 years have passed since the first report on successful genetic transformation of wheat. With the establishment and improvement of transformation platform, great progresses have been made on wheat genetic transformation both on its fundamental and applied studies in China, especially driven by the National Major Project for Transgenic Organism Breeding, China, initiated in 2008. In this review, wheat genetic transformation platform improvement and transgenic research progresses including new techniques applied and functional studies of wheat quality, yield and stress tolerant related genes and biosafety assessment are summarized. The existing problems and the trends in wheat transformation with traditional methods combined with genomic studies and genome editing technology are also discussed.展开更多
Transformation of wheat was performed by pipetting spikelets with Agrobacterium tumefaciens which contained expression vectors using Npt II as reporter gene. Transformants were identified through kana-mycin resistance...Transformation of wheat was performed by pipetting spikelets with Agrobacterium tumefaciens which contained expression vectors using Npt II as reporter gene. Transformants were identified through kana-mycin resistance, PCR and Southern blot. The results showed that transformation efficiency was within 2.0 to 3. 2% in all tested varieties of wheat. Then the simple and efficient protocol of wheat transformation by Agrobacterium tumefaciens in planta was primarily established.展开更多
INTRODUCTION There is a close relationship between the transformation of fertilizer-N and the statusof nitrogen nutrition in rhizosphere soils.Previously,Harmsen and Jager(1962)made ananalysis of N and C contents in t...INTRODUCTION There is a close relationship between the transformation of fertilizer-N and the statusof nitrogen nutrition in rhizosphere soils.Previously,Harmsen and Jager(1962)made ananalysis of N and C contents in the rhizosphere soils of upland crops and found thatclose to the root zone there was an enrichment of these substances.Recently,Qin and Liu(1984;1989)investigated the distribution of nitrogen in the rhizosphere of crops after ap-展开更多
Centromere-specific histone H3 (CENH3) replaces the canonical histone H3 in nucleosomes of functional centromeres, and plays important roles in faithful chromosome segregation during cell division. CENH3 is also impor...Centromere-specific histone H3 (CENH3) replaces the canonical histone H3 in nucleosomes of functional centromeres, and plays important roles in faithful chromosome segregation during cell division. CENH3 is also important in the recognition of alien centromeres and determines the accommodation or elimination of alien chromosomes in interspecific or intergenic hybridization. In this study, a maize full length CENH3 with a yellow fluorescent protein (YFP) tag at C-terminus (ZmCENH3-YFP) and a synthetic hybrid wmCENH3 with the N-terminus from wheat CENH3 and the histone fold domain (HFD) from maize tagged with a red fluorescent protein (RFP) at the C-terminus (wmCENH3-RFP) were transformed to wheat by biolistics transformation. Transgenic wheat plants with both ZmCNEH3-YFP and wmCENH3-RFP genes were identified by PCR. The expression of ZmCENH3-YFP was not observed, while the expression of wmCENH3-RFP could be detected by RT-PCR, direct fluorescence microscopy, and immunostaining with anti-RFP antibody. The expressed wmCENH3-RFP was localized to nuclei as dotted patterns, indicating its targeting to wheat centromeres. Somatic hybridization was performed between wmCENH3-RFP transgenic wheat and transgenic maize that expressed a ZmCENH3-YFP gene to investigate chromosome behaviors in somatic hybrids. Cytological and FISH analyses of somatic hybrid cells showed the formation of micronuclei and lagging chromatin in both somatic hybridizations with or without the wmCENH3-RFP transgene, indicating that ectopically expressed wmCENH3 could not overcome chromosome elimination in wheat/maize somatic hybrids. Immunostaining of wmCENH3-RFP and ZmCENH3-YFP in early stage somatic hybrid cells indicated that both wmCENH3-RFP and ZmCENH3-YFP proteins were expressed, but their binding patterns changed from the commonly observed dotted patterns to diffused ones, suggesting that the inactivation of CENH3 might be a factor for chromosome elimination in wheat/maize somatic hybridization.展开更多
A new approach for transforming the cultured cells of wheat(Triticum aestivum L. cv. Ganmai 8) was developed using Agrobacterium tumefaciens. The features of the optimum procedure were: (a) both combined synthetic sig...A new approach for transforming the cultured cells of wheat(Triticum aestivum L. cv. Ganmai 8) was developed using Agrobacterium tumefaciens. The features of the optimum procedure were: (a) both combined synthetic signal molecules and multiple natural extracts from susceptible plants were used to pretreat the primary vigorous Agrobacterium(PVA) cells for approximately 16 h; (b) the gyratory magnetic field condition was used during cocultivation; (c) the cocultivating period and selecting condition were modified; (d) the recipient cells were at exuberant metabolism and active division while infected with Agrobacterium. Both neomycin phosphotransferase and nopaline synthase assays demonstrated the expression of NPT II and NOS genes, located on the T-DNA segment of chimaeric plasmid pGV3850::1103neo, in transformed wheat cell colonies by adopting the techniques of dot blot, ndPAGE or high voltage paper electrophoresis. Integration of the foreign genes into wheat genome was confirmed by Southern blot hybridization. Moreover, a relatively rational method was described for the estimation of transformation frequencies from cultured cell levels.展开更多
Wheat, one of the most important food crops, has been extensively studied with respects to plant regeneration and transformation employing the immature embryos as recipient tissues. However, the transformed tissues of...Wheat, one of the most important food crops, has been extensively studied with respects to plant regeneration and transformation employing the immature embryos as recipient tissues. However, the transformed tissues often become severely necrotic after co-cultivation with Agrobacterium, which is one of the major obstacles in gene delivery. In this study, wheat varieties CB037, Kenong 199, Xinchun 9, Lunxuan 987, and Shi 4185 showed desirable culture potential or high infection ability in Agrobacterium-mediated transformation. Similarly, optimal regeneration conditions were determined by testing their ability to inhibit the cell necrosis and cell death phenotype. Two auxins of 2,4-dichlorophenoxyacetic acid (2,4-D) and 3,6-dichloro-o-anisic acid (dicamba) were evaluated for highly significant effect on both callus and plantlet production, although they were genotype-dependent in wheat. Substitution of 2,4-D by dicamba enhanced the growth and regeneration ability of callus from the immature embryos of most genotypes tested. The callus growth state couldn’t be modified by adding antioxidants such as ascorbic acid, cysteine, and silver nitrate or organic additives such as thiamine HCl and asparagine to the media. On the contrary, the best tissue statement and plant regeneration was achieved by employing the media containing the simplest MS (Murashige and Skoog) components and dicamba without organic components and vitamins. Thereby, our results are thought to inhibit wheat cell necrosis effectively and suggested to be used for more wheat genotypes.展开更多
In the protocol of wheat transformation, to use elite wheat cultivars as exogenous gene recipients can speed up the process of commercial field applications of transgenic wheat. However, it is necessary to screen whea...In the protocol of wheat transformation, to use elite wheat cultivars as exogenous gene recipients can speed up the process of commercial field applications of transgenic wheat. However, it is necessary to screen wheat cultivars with good tissue culture response (TCR) continuously from plenty of elite wheat cultivars released for wheat transformation, and it is also important to find a plant regeneration system that is suitable for these cultivars. So, the TCR of mature and immature embryos of six wheat cultivars Chuannong 11 (CN11), Chuannong12 (CN12), Chuannong17 (CN17), Chuannong18 (CN18), Chuannong19 (CN19), and Chuannong21 (CN21), which possess superior agronomic traits, were investigated by using a good TCR wheat cultivar Bobwhite as control. The results indicated that only the immature and mature embryos of CN12, CN17, and CN18 exhibited good TCR compared with Bobwhite. No significant differences were observed between embryos of Bobwhite and of the three cultivars in TCR. Mature embryo-derived calli of CN12 were used as explants for transformation by particle bombardment of SAMDC gene. Seven transformants were obtained and the efficiency was 2.3%. This research supplies three new elite recipient cultivars for wheat transformation. The wheat plant regeneration system used in this research is different from those successful ones reported previously and it could be a reference for other wheat genotypes. Furthermore, Bobwhite and the three wheat cultivars were proved to be 1RS/1BL translocation, by methods of A-PAGE, C- banding, and genomic in situ hybridization (GISH). These results imply that probably there is some relationship between 1RS/1BL translocation and TCR of wheat embryos. So this research gives us a hint that we should pay more attention to the 1RS/1BL translocations when we screen the wheat cultivars with good TCR and also that the mechanism of the effect of 1RS/ 1BL translocation on TCR is worthy of being investigated.展开更多
The specificities of tissue culture of wheat greatly limit the use of chloroplast transformation technologies in this crop. One limitation in wheat tissue culture is that it is difficult to regenerate plantlets from l...The specificities of tissue culture of wheat greatly limit the use of chloroplast transformation technologies in this crop. One limitation in wheat tissue culture is that it is difficult to regenerate plantlets from leaf tissue explants of regenerated plantlets, resulting in difficulty in obtaining homoplastic plants via multiple rounds of antibiotic selection of chloroplast transformants. Thus, a repeated in vitro regeneration system from leaf tissues was studied in this research. Our results showed that 2 mm leaf basal segments of the 4 cm high leaves from regenerated plantlets can give the best callus induction at present study. The best callus induction medium was Murashige and Skoog (MS) basal medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid and 1 mg/L naphthalenacetic acid, which gave a callus induction rate of up to 87.2%. The optimal differentiation medium was MS basal medium supplemented with 10 mg/L silver nitrate and 1 mg/L 2,3,5-triiodobenzoic acid, which gave a regeneration rate up to 33.7% for the wheat lines tested. This is the first report showing that leaf basal segments of in vitro regenerated plantlets can be used for regeneration of wheat. The establishment of a repetitive regeneration system should pave the way for the development of chloroplast transformation and the plant regeneration systems starting from leaf material of in vitro regenerated wheat and other cereal crops.展开更多
Wheat is one of the most important crops in China, and its genetic improvement has always been a research focus. In recent years, transgenic technology has been increasingly applied to improve the yield and quality, i...Wheat is one of the most important crops in China, and its genetic improvement has always been a research focus. In recent years, transgenic technology has been increasingly applied to improve the yield and quality, increase the resistance of disease, pest and stresses in wheat. The review pays particular attention to the current research progress on wheat transformation techniques, including expression vectors, reporter genes, selectable markers, genes of interest, explants and transformation methods. Based on this, the existing problems in wheat transformation techniques are discussed and the prospect of genetic breeding in wheat is also reviewed.展开更多
Wheat (Triticum aestivum L.) is a major staple food crop worldwide. It is economically important because it can be grown in a wide range of climates and geographic regions, and it has made an enormous contribution t...Wheat (Triticum aestivum L.) is a major staple food crop worldwide. It is economically important because it can be grown in a wide range of climates and geographic regions, and it has made an enormous contribution to the increase in global food production over the past four decades (Dixon et al., 2009). Wheat is produced on more than 18% of the arable land in the world, and is the most cultivated crop after maize and rice (FAOSTAT data, 2014). Despite its global strategic significance, progress in genomic and genetic engineering research on wheat has lagged behind that on other major crops due to the difficulty of culturing tissues, and the complexity of its hexaploid genome. The first successful wheat trans- formation was achieved by particle bombardment (Vasil et al., 1992). Since then additional transgenic wheat plants have been obtained by various transformation methods (Harwood, 2011). Microprojectile bombardment is considered to be a promising method, since it is robust, versatile and relatively efficient in terms of gene delivery.展开更多
Wheat ear counting is a prerequisite for the evaluation of wheat yield.A wheat ear counting method based on frequency domain decomposition is proposed in this study to improve the accuracy of wheat yield estimation.Th...Wheat ear counting is a prerequisite for the evaluation of wheat yield.A wheat ear counting method based on frequency domain decomposition is proposed in this study to improve the accuracy of wheat yield estimation.The frequency domain decomposition of wheat ear image is completed by multiscale support value filter(MSVF)combined with improved sampled contourlet transform(ISCT).Support Vector Machine(SVM)is the classic classification and regression algorithm of machine learning.MSVF based on this has strong frequency domain filtering and generalization ability,which can effectively remove the complex background,while the multi-direction characteristics of ISCT enable it to represent the contour and texture information of wheat ears.In order to improve the level of wheat yield prediction,MSVF-ISCT method is used to decompose the ear image in multiscale and multi direction in frequency domain,reduce the interference of irrelevant information,and generate the sub-band image with more abundant information components of ear feature information.Then,the ear feature is extracted by morphological operation and maximum entropy threshold segmentation,and the skeleton thinning and corner detection algorithms are used to count the results.The number of wheat ears in the image can be accurately counted.Experiments show that compared with the traditional algorithms based on spatial domain,this method significantly improves the accuracy of wheat ear counting,which can provide guidance and application for the field of agricultural precision yield estimation.展开更多
Stilbene, a kind of phytoalexin, plays an important role in resistance to fungal and bacterial infection in plants. It strongly inhibits the growth of fungi and sprout of spore. Stilbene synthase gene (Vst1) obtained ...Stilbene, a kind of phytoalexin, plays an important role in resistance to fungal and bacterial infection in plants. It strongly inhibits the growth of fungi and sprout of spore. Stilbene synthase gene (Vst1) obtained from grapevine has been transferred into common spring wheat Jinghong 5 by using the biolistic transformation method. Five transgenic plants (T0) were obtained from the bombarded 2014 immature embryos. One immune plantlet and 3 plantlets with mid-resistance to powdery mildew were identified from the transgenic plants of T3 generation which came from 2 T0 transgenic plants.展开更多
A new method was proposed to extract sensitive features and to construct a monitoring model for wheat scab based on in situ hyperspectral data of wheat ears to achieve effective prevention and control and provide theo...A new method was proposed to extract sensitive features and to construct a monitoring model for wheat scab based on in situ hyperspectral data of wheat ears to achieve effective prevention and control and provide theoretical support for its large-scale monitoring.Eight sensitive features were selected through correlation analysis and wavelet transform.These features were as follows:three original bands of 350-400 nm,500-600 nm,and 720-1000 nm;three vegetation indices of modified simple ratio(MSR),normalized difference vegetation index,and structural independent pigment index;and two wavelet features of WF01 and WF02.By combining the selected sensitive features with support vector machine(SVM)and SVM optimized by genetic algorithm(GASVM),a total of 16 monitoring models were built,and the monitoring accuracies of the two types of models were compared.The ability of the monitoring models built by GASVM to identify scab was better than that of SVM algorithm under the same characteristic variables.Among the 16 models,MSR combined with GASVM had an overall accuracy of 75%and a Kappa coefficient of 0.47.GASVM can be used to monitor wheat scab and its application can improve the accuracy of disease monitoring.展开更多
基金supported by the National Key Research and Development Program of China(2019YFC1906700)the Natural Science Foundation of Sichuan,China(2022NSFSC0308).
文摘In order to provide basic design parameters for the industrial pyrolysis process,the transformation behavior of nitrogen was investigated using wheat straw as raw material.The distributions of nitrogen in pyrolysis char,oil,and gas were obtained and the nitrogenous components in the products were analyzed systematically by X-ray photoelectron spectroscopy(XPS),pyrolysis-gas chromatography/mass spectrometry(Py-GC/MS)and thermogravimetric-Fourier transform infrared spectrometry(TG-FTIR).The nitrogen distribution ranges of the pyrolysis char,oil,and gas were 37.34%–54.82%,32.87%–40.94%and 10.20%–28.83%,respectively.More nitrogen was retained in char at lower pyrolysis temperature and the nitrogen distribution of oil was from rise to decline with increasing temperature.The most abundant N-containing compounds in three-phase products were pyrrole-N,amines,and HCN,respectively.In addition,the transformation mechanism of nitrogen from wheat straw to pyrolysis products was concluded.
基金Researchers Supporting Project Number(RSPD2024R 553),King Saud University,Riyadh,Saudi Arabia.
文摘Wheat is a critical crop,extensively consumed worldwide,and its production enhancement is essential to meet escalating demand.The presence of diseases like stem rust,leaf rust,yellow rust,and tan spot significantly diminishes wheat yield,making the early and precise identification of these diseases vital for effective disease management.With advancements in deep learning algorithms,researchers have proposed many methods for the automated detection of disease pathogens;however,accurately detectingmultiple disease pathogens simultaneously remains a challenge.This challenge arises due to the scarcity of RGB images for multiple diseases,class imbalance in existing public datasets,and the difficulty in extracting features that discriminate between multiple classes of disease pathogens.In this research,a novel method is proposed based on Transfer Generative Adversarial Networks for augmenting existing data,thereby overcoming the problems of class imbalance and data scarcity.This study proposes a customized architecture of Vision Transformers(ViT),where the feature vector is obtained by concatenating features extracted from the custom ViT and Graph Neural Networks.This paper also proposes a Model AgnosticMeta Learning(MAML)based ensemble classifier for accurate classification.The proposedmodel,validated on public datasets for wheat disease pathogen classification,achieved a test accuracy of 99.20%and an F1-score of 97.95%.Compared with existing state-of-the-art methods,this proposed model outperforms in terms of accuracy,F1-score,and the number of disease pathogens detection.In future,more diseases can be included for detection along with some other modalities like pests and weed.
基金Supported by National 863 Project (2006AA100223)Program of Introducing Talents of Discipline to Hydrology Ecological and Water Security in Arid and Semi Arid Areas(B08039)~~
文摘[Objective] The aim was to provide theoretical basis for field moisture conserving irrigation.[Method] With Xiaoyan No.6 as tested material,three different kinds of mulching irrigation treatments were carried out (straw mulching;plastic mulching;PAM control adjustment mulching).With non-mulching treatment as control,moisture conserving effect of different treatments were compared.[Result] The results showed that the water consumption of winter wheat under different soil moisture conservation treatments was low at earlier stage and later stage,but high at mid-stage,which was consistent with the water consumption law of control.There were some differences in terms of consumption intensity because of irrigation schedule and growth condition;soil moisture conservation treatments could restrain ineffective evaporation of soil moisture before anthesis.We also found that the variation of soil moisture at depth of 0-20 cm in PAM and control treatment was dramatic.The soil moisture of the former was lower than the latter at the depth of 0-20 cm,but higher at the depth of 20-50 cm.The difference of soil moisture at the depth of 0-50 cm was significant.[Conclusion] Plastic mulching and straw mulching could restrain evaporation effectively.
文摘Wheat transformation efficiency is closely related to several factors such as receptor genotype, constructed plasmid and selection procedure after bombardment or co-cultivation. In our study, several kinds of antibiotics, which were normally used in plant transformation to the selection genes of nptII, bar and hpt, were tested for the optimal concentrations for wheat transformation. The results showed that 25 - 50mg/L of geneticin (G418) was suitable for the selection of nptll, kanamycin or neomycin was not suitable for use. 3 -5mg/L of phosphinothricin (PPT) or biolaphos could be used for the selection of bar, 100 - 150mg/L of hygromycin for the selection of hpt. Yangmai 158 and Yangmai 10 with high tissue culture response and good agronomic characteristics were screened from 25 potential Chinese wheat cultivars. The concentration changing of selectable agent in selection medium was helpful to obtain enough regeneration plantlets with strong root system.
基金supported by the National Major Project for Transgenic Organism Breeding, China (2014ZX08002-004 2014ZX08010-004)+3 种基金the International S&T Cooperation Key Projects of Ministry of Science and Technology, China (2009DFB30340)the National Natural Science Foundation of China (30871524 and 31071403)the Natural Science Foundation of Hubei Province, China (2010 CBD 02403)the Wuhan Municipal S&T Research Project, China (201120922286)
文摘More than 20 years have passed since the first report on successful genetic transformation of wheat. With the establishment and improvement of transformation platform, great progresses have been made on wheat genetic transformation both on its fundamental and applied studies in China, especially driven by the National Major Project for Transgenic Organism Breeding, China, initiated in 2008. In this review, wheat genetic transformation platform improvement and transgenic research progresses including new techniques applied and functional studies of wheat quality, yield and stress tolerant related genes and biosafety assessment are summarized. The existing problems and the trends in wheat transformation with traditional methods combined with genomic studies and genome editing technology are also discussed.
基金supported by the Postdoctoral Fund of Shandong Agricultural University,China.
文摘Transformation of wheat was performed by pipetting spikelets with Agrobacterium tumefaciens which contained expression vectors using Npt II as reporter gene. Transformants were identified through kana-mycin resistance, PCR and Southern blot. The results showed that transformation efficiency was within 2.0 to 3. 2% in all tested varieties of wheat. Then the simple and efficient protocol of wheat transformation by Agrobacterium tumefaciens in planta was primarily established.
基金A part of the project supported by the National Natural Science Foundation of China
文摘INTRODUCTION There is a close relationship between the transformation of fertilizer-N and the statusof nitrogen nutrition in rhizosphere soils.Previously,Harmsen and Jager(1962)made ananalysis of N and C contents in the rhizosphere soils of upland crops and found thatclose to the root zone there was an enrichment of these substances.Recently,Qin and Liu(1984;1989)investigated the distribution of nitrogen in the rhizosphere of crops after ap-
文摘Centromere-specific histone H3 (CENH3) replaces the canonical histone H3 in nucleosomes of functional centromeres, and plays important roles in faithful chromosome segregation during cell division. CENH3 is also important in the recognition of alien centromeres and determines the accommodation or elimination of alien chromosomes in interspecific or intergenic hybridization. In this study, a maize full length CENH3 with a yellow fluorescent protein (YFP) tag at C-terminus (ZmCENH3-YFP) and a synthetic hybrid wmCENH3 with the N-terminus from wheat CENH3 and the histone fold domain (HFD) from maize tagged with a red fluorescent protein (RFP) at the C-terminus (wmCENH3-RFP) were transformed to wheat by biolistics transformation. Transgenic wheat plants with both ZmCNEH3-YFP and wmCENH3-RFP genes were identified by PCR. The expression of ZmCENH3-YFP was not observed, while the expression of wmCENH3-RFP could be detected by RT-PCR, direct fluorescence microscopy, and immunostaining with anti-RFP antibody. The expressed wmCENH3-RFP was localized to nuclei as dotted patterns, indicating its targeting to wheat centromeres. Somatic hybridization was performed between wmCENH3-RFP transgenic wheat and transgenic maize that expressed a ZmCENH3-YFP gene to investigate chromosome behaviors in somatic hybrids. Cytological and FISH analyses of somatic hybrid cells showed the formation of micronuclei and lagging chromatin in both somatic hybridizations with or without the wmCENH3-RFP transgene, indicating that ectopically expressed wmCENH3 could not overcome chromosome elimination in wheat/maize somatic hybrids. Immunostaining of wmCENH3-RFP and ZmCENH3-YFP in early stage somatic hybrid cells indicated that both wmCENH3-RFP and ZmCENH3-YFP proteins were expressed, but their binding patterns changed from the commonly observed dotted patterns to diffused ones, suggesting that the inactivation of CENH3 might be a factor for chromosome elimination in wheat/maize somatic hybridization.
文摘A new approach for transforming the cultured cells of wheat(Triticum aestivum L. cv. Ganmai 8) was developed using Agrobacterium tumefaciens. The features of the optimum procedure were: (a) both combined synthetic signal molecules and multiple natural extracts from susceptible plants were used to pretreat the primary vigorous Agrobacterium(PVA) cells for approximately 16 h; (b) the gyratory magnetic field condition was used during cocultivation; (c) the cocultivating period and selecting condition were modified; (d) the recipient cells were at exuberant metabolism and active division while infected with Agrobacterium. Both neomycin phosphotransferase and nopaline synthase assays demonstrated the expression of NPT II and NOS genes, located on the T-DNA segment of chimaeric plasmid pGV3850::1103neo, in transformed wheat cell colonies by adopting the techniques of dot blot, ndPAGE or high voltage paper electrophoresis. Integration of the foreign genes into wheat genome was confirmed by Southern blot hybridization. Moreover, a relatively rational method was described for the estimation of transformation frequencies from cultured cell levels.
基金supported by the National Natural Science Foundation of China (30971776)the National Transgenic Specialized Research Program of China (2008ZX08010-004)
文摘Wheat, one of the most important food crops, has been extensively studied with respects to plant regeneration and transformation employing the immature embryos as recipient tissues. However, the transformed tissues often become severely necrotic after co-cultivation with Agrobacterium, which is one of the major obstacles in gene delivery. In this study, wheat varieties CB037, Kenong 199, Xinchun 9, Lunxuan 987, and Shi 4185 showed desirable culture potential or high infection ability in Agrobacterium-mediated transformation. Similarly, optimal regeneration conditions were determined by testing their ability to inhibit the cell necrosis and cell death phenotype. Two auxins of 2,4-dichlorophenoxyacetic acid (2,4-D) and 3,6-dichloro-o-anisic acid (dicamba) were evaluated for highly significant effect on both callus and plantlet production, although they were genotype-dependent in wheat. Substitution of 2,4-D by dicamba enhanced the growth and regeneration ability of callus from the immature embryos of most genotypes tested. The callus growth state couldn’t be modified by adding antioxidants such as ascorbic acid, cysteine, and silver nitrate or organic additives such as thiamine HCl and asparagine to the media. On the contrary, the best tissue statement and plant regeneration was achieved by employing the media containing the simplest MS (Murashige and Skoog) components and dicamba without organic components and vitamins. Thereby, our results are thought to inhibit wheat cell necrosis effectively and suggested to be used for more wheat genotypes.
基金We thank Dr.Yang Zujun,Zhang Huaiyu,Yan Benju,Tan Feiquan,and Zhou Jianpin for their helpful comments in improving the manuscript.We also thank Cheng Jing for providing wheat cv.Bobwhite.This work was supported by 948 Project of Ministry of Agriculture,China(246)the National Natural Science Foundation of China(30170579).
文摘In the protocol of wheat transformation, to use elite wheat cultivars as exogenous gene recipients can speed up the process of commercial field applications of transgenic wheat. However, it is necessary to screen wheat cultivars with good tissue culture response (TCR) continuously from plenty of elite wheat cultivars released for wheat transformation, and it is also important to find a plant regeneration system that is suitable for these cultivars. So, the TCR of mature and immature embryos of six wheat cultivars Chuannong 11 (CN11), Chuannong12 (CN12), Chuannong17 (CN17), Chuannong18 (CN18), Chuannong19 (CN19), and Chuannong21 (CN21), which possess superior agronomic traits, were investigated by using a good TCR wheat cultivar Bobwhite as control. The results indicated that only the immature and mature embryos of CN12, CN17, and CN18 exhibited good TCR compared with Bobwhite. No significant differences were observed between embryos of Bobwhite and of the three cultivars in TCR. Mature embryo-derived calli of CN12 were used as explants for transformation by particle bombardment of SAMDC gene. Seven transformants were obtained and the efficiency was 2.3%. This research supplies three new elite recipient cultivars for wheat transformation. The wheat plant regeneration system used in this research is different from those successful ones reported previously and it could be a reference for other wheat genotypes. Furthermore, Bobwhite and the three wheat cultivars were proved to be 1RS/1BL translocation, by methods of A-PAGE, C- banding, and genomic in situ hybridization (GISH). These results imply that probably there is some relationship between 1RS/1BL translocation and TCR of wheat embryos. So this research gives us a hint that we should pay more attention to the 1RS/1BL translocations when we screen the wheat cultivars with good TCR and also that the mechanism of the effect of 1RS/ 1BL translocation on TCR is worthy of being investigated.
文摘The specificities of tissue culture of wheat greatly limit the use of chloroplast transformation technologies in this crop. One limitation in wheat tissue culture is that it is difficult to regenerate plantlets from leaf tissue explants of regenerated plantlets, resulting in difficulty in obtaining homoplastic plants via multiple rounds of antibiotic selection of chloroplast transformants. Thus, a repeated in vitro regeneration system from leaf tissues was studied in this research. Our results showed that 2 mm leaf basal segments of the 4 cm high leaves from regenerated plantlets can give the best callus induction at present study. The best callus induction medium was Murashige and Skoog (MS) basal medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid and 1 mg/L naphthalenacetic acid, which gave a callus induction rate of up to 87.2%. The optimal differentiation medium was MS basal medium supplemented with 10 mg/L silver nitrate and 1 mg/L 2,3,5-triiodobenzoic acid, which gave a regeneration rate up to 33.7% for the wheat lines tested. This is the first report showing that leaf basal segments of in vitro regenerated plantlets can be used for regeneration of wheat. The establishment of a repetitive regeneration system should pave the way for the development of chloroplast transformation and the plant regeneration systems starting from leaf material of in vitro regenerated wheat and other cereal crops.
基金Supported by the Basic and Frontier Technology Research Programs from the Department of Science and Technology of Henan Province (072300430120)
文摘Wheat is one of the most important crops in China, and its genetic improvement has always been a research focus. In recent years, transgenic technology has been increasingly applied to improve the yield and quality, increase the resistance of disease, pest and stresses in wheat. The review pays particular attention to the current research progress on wheat transformation techniques, including expression vectors, reporter genes, selectable markers, genes of interest, explants and transformation methods. Based on this, the existing problems in wheat transformation techniques are discussed and the prospect of genetic breeding in wheat is also reviewed.
基金funded by the Ministry of Agriculture of China(Nos.2014ZX0801003B and 2013ZX08002-004)
文摘Wheat (Triticum aestivum L.) is a major staple food crop worldwide. It is economically important because it can be grown in a wide range of climates and geographic regions, and it has made an enormous contribution to the increase in global food production over the past four decades (Dixon et al., 2009). Wheat is produced on more than 18% of the arable land in the world, and is the most cultivated crop after maize and rice (FAOSTAT data, 2014). Despite its global strategic significance, progress in genomic and genetic engineering research on wheat has lagged behind that on other major crops due to the difficulty of culturing tissues, and the complexity of its hexaploid genome. The first successful wheat trans- formation was achieved by particle bombardment (Vasil et al., 1992). Since then additional transgenic wheat plants have been obtained by various transformation methods (Harwood, 2011). Microprojectile bombardment is considered to be a promising method, since it is robust, versatile and relatively efficient in terms of gene delivery.
基金National Natural Science Foundation of China(61672032)National Key Research and Development Program of China(2016YFD0800904)+1 种基金Anhui Provincial Science and Technology Project(16030701091)The Open Research Fund of National Engineering Research Center for Agro-Ecological Big Data Analysis&Application,Anhui University(AE2018009).
文摘Wheat ear counting is a prerequisite for the evaluation of wheat yield.A wheat ear counting method based on frequency domain decomposition is proposed in this study to improve the accuracy of wheat yield estimation.The frequency domain decomposition of wheat ear image is completed by multiscale support value filter(MSVF)combined with improved sampled contourlet transform(ISCT).Support Vector Machine(SVM)is the classic classification and regression algorithm of machine learning.MSVF based on this has strong frequency domain filtering and generalization ability,which can effectively remove the complex background,while the multi-direction characteristics of ISCT enable it to represent the contour and texture information of wheat ears.In order to improve the level of wheat yield prediction,MSVF-ISCT method is used to decompose the ear image in multiscale and multi direction in frequency domain,reduce the interference of irrelevant information,and generate the sub-band image with more abundant information components of ear feature information.Then,the ear feature is extracted by morphological operation and maximum entropy threshold segmentation,and the skeleton thinning and corner detection algorithms are used to count the results.The number of wheat ears in the image can be accurately counted.Experiments show that compared with the traditional algorithms based on spatial domain,this method significantly improves the accuracy of wheat ear counting,which can provide guidance and application for the field of agricultural precision yield estimation.
文摘Stilbene, a kind of phytoalexin, plays an important role in resistance to fungal and bacterial infection in plants. It strongly inhibits the growth of fungi and sprout of spore. Stilbene synthase gene (Vst1) obtained from grapevine has been transferred into common spring wheat Jinghong 5 by using the biolistic transformation method. Five transgenic plants (T0) were obtained from the bombarded 2014 immature embryos. One immune plantlet and 3 plantlets with mid-resistance to powdery mildew were identified from the transgenic plants of T3 generation which came from 2 T0 transgenic plants.
基金This work was supported by National Natural Science Foundation of China(41571354,41871339)Natural Science Research Project of Anhui Provincial Education Department(KJ2019A0030)+2 种基金Anhui Provincial Science and Technology Project(16030701091,201904f06020038)supported by Hainan Provincial Key R&D Program of China(ZDYF2018073)National special support program for high-level personnel recruitment(Wenjiang Huang).
文摘A new method was proposed to extract sensitive features and to construct a monitoring model for wheat scab based on in situ hyperspectral data of wheat ears to achieve effective prevention and control and provide theoretical support for its large-scale monitoring.Eight sensitive features were selected through correlation analysis and wavelet transform.These features were as follows:three original bands of 350-400 nm,500-600 nm,and 720-1000 nm;three vegetation indices of modified simple ratio(MSR),normalized difference vegetation index,and structural independent pigment index;and two wavelet features of WF01 and WF02.By combining the selected sensitive features with support vector machine(SVM)and SVM optimized by genetic algorithm(GASVM),a total of 16 monitoring models were built,and the monitoring accuracies of the two types of models were compared.The ability of the monitoring models built by GASVM to identify scab was better than that of SVM algorithm under the same characteristic variables.Among the 16 models,MSR combined with GASVM had an overall accuracy of 75%and a Kappa coefficient of 0.47.GASVM can be used to monitor wheat scab and its application can improve the accuracy of disease monitoring.