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
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
Electroporation, PEG, PEG plus electroporation and Biolistics methods were tested ingene transformation of P. yezoensis. The exogenous gus was from plasmid of pBI121 and pCAMBI-A1301, both contain the CaMV35S promoter...Electroporation, PEG, PEG plus electroporation and Biolistics methods were tested ingene transformation of P. yezoensis. The exogenous gus was from plasmid of pBI121 and pCAMBI-A1301, both contain the CaMV35S promoter. The receptors included the prooplasts, tissues and free-liv-ing conchocelis filaments of P. yezoensis. Several factors, for example, the voltage, capacitance and bi-valent cations, etc., were studied. Results show that these four methods are all efficient for gene transfor-mation in P. yezoensis; and that PEG is the best one, with transformation efficiency of up to 4 ×10-5.GUS activity was detected 26 days after transfation by using PEG method.展开更多
Cedrela odorata is a tropical tree widely appreciated for its wood. Commercial plantations are frequently hampered by the attack of the meliacea borer, Hypsipyla grandella, and the lack of resistant varieties. C. odor...Cedrela odorata is a tropical tree widely appreciated for its wood. Commercial plantations are frequently hampered by the attack of the meliacea borer, Hypsipyla grandella, and the lack of resistant varieties. C. odorata traditional breeding would consume very long periods of time, thus direct transfer of entomotoxic coding genes to generate resistant varieties is a promising alternative. There are two prerequisites for gene manipulation of this species: 1) to set the conditions for transgene delivery and 2) to have a way to select regenerating transformed plants. In this paper, we report the optimal biolistics conditions for transient expression of uidA and gfp reporter genes in C. odorata somatic embryos and the selective doses for kanamycin, spectinomycin, phosphinotrycin and hygromycin to screen transformed cells.展开更多
2-3 anti-fungal disease genes are coinserted with hygromycin phosphotransferase in the same vector. Two insecticidal genes and PPT acetyl transferase genes are placed in another one. The vectors are co-delivered to ri...2-3 anti-fungal disease genes are coinserted with hygromycin phosphotransferase in the same vector. Two insecticidal genes and PPT acetyl transferase genes are placed in another one. The vectors are co-delivered to rice embryonic cellus tissue at a molar ratio of 1:1 using the particle gun method. 55 independent regenerated lines have been obtained through screening for hygromycin resistance. Of these, 70% transgenic plants harbor 6-7 foreign genes. The genes on the same vectors are always co-delivered to rice plant. Northern blot analysis has indicated that the multiple foreign genes give stable expression. In the 6 transgenic plants carrying 6-7 foreign genes, multiple foreign genes tend to integrate in 1 or 2 genetic loci. Progeny segregation is consistent with Mendel’s 3:1 segregation law. 8 homozy-gous R1 transgenic plants harboring 2-3 anti-fungal and 2 insecticidal genes are selected from large number of transgenic progeny screening for hygromycin and Basta resistance.展开更多
A senescence-inhibition chimeric gene containing the specific promoter of SAG 12 and IPT gene was transferred into rice with the biolistic method. Results of PCR, Dot blotting and Southern blotting indicated that the ...A senescence-inhibition chimeric gene containing the specific promoter of SAG 12 and IPT gene was transferred into rice with the biolistic method. Results of PCR, Dot blotting and Southern blotting indicated that the chimeric gene had been integrated into rice genome. Analyses of GUS activity and cytokinin content in transgenic plants of rice and the observation of T 1 generation plant at grain formation stage indicated that the foreign gene was expressed.展开更多
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.
文摘Electroporation, PEG, PEG plus electroporation and Biolistics methods were tested ingene transformation of P. yezoensis. The exogenous gus was from plasmid of pBI121 and pCAMBI-A1301, both contain the CaMV35S promoter. The receptors included the prooplasts, tissues and free-liv-ing conchocelis filaments of P. yezoensis. Several factors, for example, the voltage, capacitance and bi-valent cations, etc., were studied. Results show that these four methods are all efficient for gene transfor-mation in P. yezoensis; and that PEG is the best one, with transformation efficiency of up to 4 ×10-5.GUS activity was detected 26 days after transfation by using PEG method.
基金support provided by CONACYT-CONA-FOR C03-10013SEP-CONACYT C01-53851ITSA-DIC 2004-1
文摘Cedrela odorata is a tropical tree widely appreciated for its wood. Commercial plantations are frequently hampered by the attack of the meliacea borer, Hypsipyla grandella, and the lack of resistant varieties. C. odorata traditional breeding would consume very long periods of time, thus direct transfer of entomotoxic coding genes to generate resistant varieties is a promising alternative. There are two prerequisites for gene manipulation of this species: 1) to set the conditions for transgene delivery and 2) to have a way to select regenerating transformed plants. In this paper, we report the optimal biolistics conditions for transient expression of uidA and gfp reporter genes in C. odorata somatic embryos and the selective doses for kanamycin, spectinomycin, phosphinotrycin and hygromycin to screen transformed cells.
基金the State "863" High-Tech Project and the specific fund for rice of The South China Life Science and Technology Research Center from The National Science & Technology Department.
文摘2-3 anti-fungal disease genes are coinserted with hygromycin phosphotransferase in the same vector. Two insecticidal genes and PPT acetyl transferase genes are placed in another one. The vectors are co-delivered to rice embryonic cellus tissue at a molar ratio of 1:1 using the particle gun method. 55 independent regenerated lines have been obtained through screening for hygromycin resistance. Of these, 70% transgenic plants harbor 6-7 foreign genes. The genes on the same vectors are always co-delivered to rice plant. Northern blot analysis has indicated that the multiple foreign genes give stable expression. In the 6 transgenic plants carrying 6-7 foreign genes, multiple foreign genes tend to integrate in 1 or 2 genetic loci. Progeny segregation is consistent with Mendel’s 3:1 segregation law. 8 homozy-gous R1 transgenic plants harboring 2-3 anti-fungal and 2 insecticidal genes are selected from large number of transgenic progeny screening for hygromycin and Basta resistance.
文摘A senescence-inhibition chimeric gene containing the specific promoter of SAG 12 and IPT gene was transferred into rice with the biolistic method. Results of PCR, Dot blotting and Southern blotting indicated that the chimeric gene had been integrated into rice genome. Analyses of GUS activity and cytokinin content in transgenic plants of rice and the observation of T 1 generation plant at grain formation stage indicated that the foreign gene was expressed.