Wheat, triticale, tritordeum, barley, oat and rye are the most important crops in human consumptions and industry in the world. Transformation technology supplies a new source of improving Triticeae crops. In the past...Wheat, triticale, tritordeum, barley, oat and rye are the most important crops in human consumptions and industry in the world. Transformation technology supplies a new source of improving Triticeae crops. In the past decade, transformation of wheat crops has considerably progressed. Many transgenic plants of Triticeae crops with various genes were produced via nricroprojectile bombardment, Agrobacterium-mediated transformation, PEG-uptake DNA technique, electroporation, microinjection, injection inflorescence and silicone carbide. Integration and expression of transgenes, inheritance and variation of transgenic plants have been studied. Technical improvements of genetic transformation for wheat crops will be extensively useful in commerce and benefit significantly to human being in the world.展开更多
The soft rot infected by pathogenic bacterium Erwinia aroideae Holland is one of the three serious diseases of Chinese cabbage ( Brassica pekinensis Rupr.). By constructing vector system of high frequency transformati...The soft rot infected by pathogenic bacterium Erwinia aroideae Holland is one of the three serious diseases of Chinese cabbage ( Brassica pekinensis Rupr.). By constructing vector system of high frequency transformation mediated by Agrobacterium tunefaciens EHA105, anti-bacterial peptide gene with strong bactericidal action to pathogenic bacteria was introduced into Chinese cabbage AB-81 self-bred line and the transgenic plants were obtained. PCR and Southern blotting detection showed that target gene was integrated into plant genome of Chinese cabbage. The tests of bacteriostasis action of the extract from transgenic plants in vitro, and the assay of disease-resistant of transgenic plantlets in the tube and the pot by perfusing inoculation with pathogenic bacteria showed obvious resistance to soft rot. This resistance can be a stable heredity by genetic analysis of generations of transgenic plants self-bred, separation ratio of its R, was 3:1. The resistance to Km and disease of soft rot was still kept in the R-5. These results indicated the possibility of breeding new varieties of anti-soft rot Chinese cabbage by transgenic plants as parents.展开更多
[Objective] This research aimed to solve the problems of growth and differentiation inhibition of transgenic potato plants caused by antibiotics used for bacteriostasis. [Method] Microtubers were induced using transge...[Objective] This research aimed to solve the problems of growth and differentiation inhibition of transgenic potato plants caused by antibiotics used for bacteriostasis. [Method] Microtubers were induced using transgenic potato plants, which had generated shoots and formed transgenic bacteria-free plants. [Result] Among the three transgenic potato varieties, the optimal induction medium for SⅠ and SⅡ were MS＋ 0.5 mg/L of 6-BA ＋ 0.1 mg/L of GA3＋ 150 mg/L of cef, and the optimal induction medium for NT were MS＋ 0.5 mg/L of ZT ＋ 0.1 mg/L of GA3 ＋ 150 mg/L of cef; the optimal differentiation medium for tubers were MS＋ 0.5 mg/L of ZT ＋ 0.1 mg/L of NAA, and the tubers with diameters ranging from 0.5 to 0.7 cm had generated the most shoots. The transgenic bacteria-free plants were cultivated in propagation medium without antibiotics for 30 d with a contamination rate of 0, and the stems of bacteria-free plants were stout with no branching. [Conclusion] This method is simple and could be easily applied for the removal of bacteria, which had cleared away obstacles for the selection and growth of transgenic individuals.展开更多
Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, 'Xinluzao_1', 'Jinmian_7', 'Jinmian_12' and 'Jihe_321&#...Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, 'Xinluzao_1', 'Jinmian_7', 'Jinmian_12' and 'Jihe_321' were transformed respectively by two efficient plant expression plasmids pBinMoBc and pBinoBc via Agrobacterium tumefaciens . In pBinMoBc, cry 1Ac3 gene, which encodes the Bt toxin, is under the control of chimeric OM promoter. In pBinoBc, it is under control of CaMV 35S promoter. After co_cultivation with Agrobacterium tumefimpfaciens LBA4404 (containing pBinMoBc or pBinoBc), kanamycin_resistant selection, somatic embryos were induced and regenerated plants were obtained. Then the regenerated plantlets were grafted to untransformed stocks in greenhouse to produce descendants. The integration of cry 1Ac3 gene and its expression in T 2 generation of transgenic cotton plants were confirmed by Southern hybridization and Western blotting. The analyses of insect bioassay indicated that the transgenic plants of both constructions have significant resistance to the larvae of cotton bollworm ( Heliothis armigera ) and that cry 1Ac3 gene driven by chimeric OM promoter could endue T 2 generation cotton with high pest_resistant ability, implicating that it has a profound application in genetic engineering to breed new pest_resistant cotton varieties.展开更多
Chloroplast genetic engineering, with several advantages over nuclear genetic engineering, is now regarded as an attractive new technology in basic and applied research, including deepening our understanding of plasti...Chloroplast genetic engineering, with several advantages over nuclear genetic engineering, is now regarded as an attractive new technology in basic and applied research, including deepening our understanding of plastid genome, engineering plant metabolic system, generating transplastomic plants with higher resistance to insect, disease, drought and herbicide and bioproducing of antibodies and vaccines. In this review, the principle and operating system for chloroplast genetic engineering and its application in higher plants have been discussed.展开更多
The experiment was carried out on five different species of Paulownia for callus induction from leaves. MS medium was adopted as basic medium, and from different combinations of NAA and BA the suitable media were dete...The experiment was carried out on five different species of Paulownia for callus induction from leaves. MS medium was adopted as basic medium, and from different combinations of NAA and BA the suitable media were determined for callus induction, bud differentiation, and root differentiation of five different species. MS+0.5NAA+4BA, MS+0.3NAA+2BA, MS+0.5NAA+4BA, MS+0.3NAA+6BA, and MS+0.3NAA+8BA were suitable media of callus inductions of leaves, respectively, for Paulownia tomentosa, Paulownia australis, Paulownia fortunei, Paulownia elongata and P. tmentosa x P. fortunei, and MS+0.3NAA+12BA, MS+0.3NAA+12BA, MS+0.5NAA+12BA, MS+0.5NAA+12BA, and MS+0.7NAA+12BA were suitable media for bud differentiation from leaf callus respectively for above five species. The rooting media was determined as 2MS+0.1NAA, 1/2MS+0.1NAA, 1/2MS, 1/2MS+0.3NAA, and 1/2MS+0.5NAA. These results provide reference data for breeding new fine va-rieties with different kinds of Paulownia protoplasts fusions.展开更多
The success for genetic transformation of maize (Zea mays L.) is highly related to genotype of target material. A few model varieties can be induced into type Ⅱ callus, which can be easily transformed with high regen...The success for genetic transformation of maize (Zea mays L.) is highly related to genotype of target material. A few model varieties can be induced into type Ⅱ callus, which can be easily transformed with high regeneration frequency. However, most of cultivars could be only induced into type Ⅰ callus, which is difficult to be transformed with low regeneration. Thus, studying on the conditions of induction and transformation for type Ⅰ callus will show great importance for improving elite of maize directly with genetic engineering. Bacillus thuringiensis toxin protein (cry1Ac3) gene was successfully delivered into type Ⅰ calli of two elite inbred lines of maize, 340 and E28, via particle bombardment in this work. Fertile transgenic corn plants were obtained through phosphinothricin (PPT) or hygromycin B (HygB) selection, and the results of PCR, Southern blot assay and ELISA showed that foreign genes had been integrated into maize genome and expressed. In the meantime, strong resistance of some transgenic plants to corn borer was showed through bioassay. In addition, the comparison of selective effect between PPT and HygB showed that PPT, as a selective agent, was better than HygB for the growth and regeneration of resistant calli.展开更多
[Objective]The research aimed to study the regeneration technology of Cazania rigens L.leaves and screen out the optimum medium formula for the regeneration of Cazania rigens L.leaves.[Method]Using Japan imported C.ri...[Objective]The research aimed to study the regeneration technology of Cazania rigens L.leaves and screen out the optimum medium formula for the regeneration of Cazania rigens L.leaves.[Method]Using Japan imported C.rigens leaves as materials,the orthogonal test was made for the callus and adventitious buds induction in MS medium with different kinds and concentrations of hormones.The optimum medium formula for the regeneration of C.rigens leaves were screened out.[Result]On the medium of MS ＋ 0.8-1.0 mg/L TDZ ＋ 0.05-1.0 mg/L NAA,compact type and bright green calli were formed.When the leaves were inoculated on the medium of MS ＋ 0.5-1.0 mg/L TDZ ＋ 0.05-1.0 mg/L NAA,many adventitious shoots can be induced and the induction rate reached 100%.When strong adventitious shoots with the height of 2.0-3.0 cm were transplanted into the medium of 1/2 MS ＋0.1 mg/L NAA,the rooting situations were good and the rooting rate was 100%.[Conclusion]The research provided a new way for the rapid propagation of C.rigens and laid the foundation for the genetic transformation and new varieties breeding of C.rigens.展开更多
文摘Wheat, triticale, tritordeum, barley, oat and rye are the most important crops in human consumptions and industry in the world. Transformation technology supplies a new source of improving Triticeae crops. In the past decade, transformation of wheat crops has considerably progressed. Many transgenic plants of Triticeae crops with various genes were produced via nricroprojectile bombardment, Agrobacterium-mediated transformation, PEG-uptake DNA technique, electroporation, microinjection, injection inflorescence and silicone carbide. Integration and expression of transgenes, inheritance and variation of transgenic plants have been studied. Technical improvements of genetic transformation for wheat crops will be extensively useful in commerce and benefit significantly to human being in the world.
文摘The soft rot infected by pathogenic bacterium Erwinia aroideae Holland is one of the three serious diseases of Chinese cabbage ( Brassica pekinensis Rupr.). By constructing vector system of high frequency transformation mediated by Agrobacterium tunefaciens EHA105, anti-bacterial peptide gene with strong bactericidal action to pathogenic bacteria was introduced into Chinese cabbage AB-81 self-bred line and the transgenic plants were obtained. PCR and Southern blotting detection showed that target gene was integrated into plant genome of Chinese cabbage. The tests of bacteriostasis action of the extract from transgenic plants in vitro, and the assay of disease-resistant of transgenic plantlets in the tube and the pot by perfusing inoculation with pathogenic bacteria showed obvious resistance to soft rot. This resistance can be a stable heredity by genetic analysis of generations of transgenic plants self-bred, separation ratio of its R, was 3:1. The resistance to Km and disease of soft rot was still kept in the R-5. These results indicated the possibility of breeding new varieties of anti-soft rot Chinese cabbage by transgenic plants as parents.
基金Supported by the Open Project of National Engineering Laboratory for Tree Breeding of Beijing Forestry University~~
文摘[Objective] This research aimed to solve the problems of growth and differentiation inhibition of transgenic potato plants caused by antibiotics used for bacteriostasis. [Method] Microtubers were induced using transgenic potato plants, which had generated shoots and formed transgenic bacteria-free plants. [Result] Among the three transgenic potato varieties, the optimal induction medium for SⅠ and SⅡ were MS＋ 0.5 mg/L of 6-BA ＋ 0.1 mg/L of GA3＋ 150 mg/L of cef, and the optimal induction medium for NT were MS＋ 0.5 mg/L of ZT ＋ 0.1 mg/L of GA3 ＋ 150 mg/L of cef; the optimal differentiation medium for tubers were MS＋ 0.5 mg/L of ZT ＋ 0.1 mg/L of NAA, and the tubers with diameters ranging from 0.5 to 0.7 cm had generated the most shoots. The transgenic bacteria-free plants were cultivated in propagation medium without antibiotics for 30 d with a contamination rate of 0, and the stems of bacteria-free plants were stout with no branching. [Conclusion] This method is simple and could be easily applied for the removal of bacteria, which had cleared away obstacles for the selection and growth of transgenic individuals.
文摘Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, 'Xinluzao_1', 'Jinmian_7', 'Jinmian_12' and 'Jihe_321' were transformed respectively by two efficient plant expression plasmids pBinMoBc and pBinoBc via Agrobacterium tumefaciens . In pBinMoBc, cry 1Ac3 gene, which encodes the Bt toxin, is under the control of chimeric OM promoter. In pBinoBc, it is under control of CaMV 35S promoter. After co_cultivation with Agrobacterium tumefimpfaciens LBA4404 (containing pBinMoBc or pBinoBc), kanamycin_resistant selection, somatic embryos were induced and regenerated plants were obtained. Then the regenerated plantlets were grafted to untransformed stocks in greenhouse to produce descendants. The integration of cry 1Ac3 gene and its expression in T 2 generation of transgenic cotton plants were confirmed by Southern hybridization and Western blotting. The analyses of insect bioassay indicated that the transgenic plants of both constructions have significant resistance to the larvae of cotton bollworm ( Heliothis armigera ) and that cry 1Ac3 gene driven by chimeric OM promoter could endue T 2 generation cotton with high pest_resistant ability, implicating that it has a profound application in genetic engineering to breed new pest_resistant cotton varieties.
文摘Chloroplast genetic engineering, with several advantages over nuclear genetic engineering, is now regarded as an attractive new technology in basic and applied research, including deepening our understanding of plastid genome, engineering plant metabolic system, generating transplastomic plants with higher resistance to insect, disease, drought and herbicide and bioproducing of antibodies and vaccines. In this review, the principle and operating system for chloroplast genetic engineering and its application in higher plants have been discussed.
基金This paper was supported by National Nature Science Foundation of China (No. 39870631) and Nature Science Foundation of Henan Province (No. 994011100).
文摘The experiment was carried out on five different species of Paulownia for callus induction from leaves. MS medium was adopted as basic medium, and from different combinations of NAA and BA the suitable media were determined for callus induction, bud differentiation, and root differentiation of five different species. MS+0.5NAA+4BA, MS+0.3NAA+2BA, MS+0.5NAA+4BA, MS+0.3NAA+6BA, and MS+0.3NAA+8BA were suitable media of callus inductions of leaves, respectively, for Paulownia tomentosa, Paulownia australis, Paulownia fortunei, Paulownia elongata and P. tmentosa x P. fortunei, and MS+0.3NAA+12BA, MS+0.3NAA+12BA, MS+0.5NAA+12BA, MS+0.5NAA+12BA, and MS+0.7NAA+12BA were suitable media for bud differentiation from leaf callus respectively for above five species. The rooting media was determined as 2MS+0.1NAA, 1/2MS+0.1NAA, 1/2MS, 1/2MS+0.3NAA, and 1/2MS+0.5NAA. These results provide reference data for breeding new fine va-rieties with different kinds of Paulownia protoplasts fusions.
文摘The success for genetic transformation of maize (Zea mays L.) is highly related to genotype of target material. A few model varieties can be induced into type Ⅱ callus, which can be easily transformed with high regeneration frequency. However, most of cultivars could be only induced into type Ⅰ callus, which is difficult to be transformed with low regeneration. Thus, studying on the conditions of induction and transformation for type Ⅰ callus will show great importance for improving elite of maize directly with genetic engineering. Bacillus thuringiensis toxin protein (cry1Ac3) gene was successfully delivered into type Ⅰ calli of two elite inbred lines of maize, 340 and E28, via particle bombardment in this work. Fertile transgenic corn plants were obtained through phosphinothricin (PPT) or hygromycin B (HygB) selection, and the results of PCR, Southern blot assay and ELISA showed that foreign genes had been integrated into maize genome and expressed. In the meantime, strong resistance of some transgenic plants to corn borer was showed through bioassay. In addition, the comparison of selective effect between PPT and HygB showed that PPT, as a selective agent, was better than HygB for the growth and regeneration of resistant calli.
基金Supported by the Extra-curricular Academic Research Found of 12th Batch of Students in Soochow University (KY2010114A)Science and Technology Support (Agriculture) Project of Suzhou Province(SNG0908)~~
文摘[Objective]The research aimed to study the regeneration technology of Cazania rigens L.leaves and screen out the optimum medium formula for the regeneration of Cazania rigens L.leaves.[Method]Using Japan imported C.rigens leaves as materials,the orthogonal test was made for the callus and adventitious buds induction in MS medium with different kinds and concentrations of hormones.The optimum medium formula for the regeneration of C.rigens leaves were screened out.[Result]On the medium of MS ＋ 0.8-1.0 mg/L TDZ ＋ 0.05-1.0 mg/L NAA,compact type and bright green calli were formed.When the leaves were inoculated on the medium of MS ＋ 0.5-1.0 mg/L TDZ ＋ 0.05-1.0 mg/L NAA,many adventitious shoots can be induced and the induction rate reached 100%.When strong adventitious shoots with the height of 2.0-3.0 cm were transplanted into the medium of 1/2 MS ＋0.1 mg/L NAA,the rooting situations were good and the rooting rate was 100%.[Conclusion]The research provided a new way for the rapid propagation of C.rigens and laid the foundation for the genetic transformation and new varieties breeding of C.rigens.
