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
The genetic transformation was achieved by pollen_mediated approach on maize ( Zea mays L.) inbred lines Tai 9101 and Zong 31. Plasmid DNA of pGLⅡ_RC_1 was mixed with fresh pollen of maize inbreds in sucrose ...The genetic transformation was achieved by pollen_mediated approach on maize ( Zea mays L.) inbred lines Tai 9101 and Zong 31. Plasmid DNA of pGLⅡ_RC_1 was mixed with fresh pollen of maize inbreds in sucrose solution. The pollens were treated by ultrasonication and collected, pollinated on silks of maize ears. Transformants were confirmed by dot blot hybridization, PCR amplification and PCR_Southern blot hybridization. The pollen_mediated transformation approach could circumvent the tedious tissue culture procedures like in particle bombardment and Agrobacterium infection, etc. This approach is simple, easy to operate, and could be widely used in practice.展开更多
A 9-week feeding trial in floating freshwater cages(1.0 m×1.0 m×2.0 m) was conducted to study the effects of different dietary levels of protein and starch on growth,body composition,and gene expression of e...A 9-week feeding trial in floating freshwater cages(1.0 m×1.0 m×2.0 m) was conducted to study the effects of different dietary levels of protein and starch on growth,body composition,and gene expression of enzymes in common carp,Cyprinus carpio(mean body weight,36.12±1.18 g) to evaluate the protein-sparing effect of dietary carbohydrate.Four diets were formulated with corn starch as the carbohydrate source to obtain corn starch levels of 6.5%,13%,19.5%,or 26%and protein levels of 30.5%,28.2%,26.4%,and 24.2%.The results showed no differences in growth performance of fish fed the diets with different protein and corn starch levels,but body composition and glucose metabolic enzyme activity of carp were significantly affected by the different diets(P<0.05).Weight gain,specific growth rate,and the feed conversion ratio were not different in fish fed the different dietary treatments.Protein efficiency ratio increased significantly as corn starch level increased(P<0.05).Whole-body crude lipid composition increased with increasing dietary corn starch level(P<0.05).Glucokinase(GK),hexokinase,and pyruvate kinase(PK) activities increased significantly with increasing dietary corn starch level(P<0.05),whereas glucose-6-phosphate(G6Pase) activity decreased with increasing dietary corn starch level(P<0.05).GK gene expression was significantly higher in fish fed the high-corn starch diet than those fed the low-corn starch diet(P<0.05).G6 pase gene expression tended to decrease with increasing starch level(P>0.05).In summary,the results indicate a protein-sparing effect by substituting carbohydrate in the diet of common carp.展开更多
Five new compounds were tested on the growth and antifeeding activity compared with toosendanin against fifth instar larvae Ostrinia furnacalis. The activities of two proteases, a weak alkaline trypsine like enzy...Five new compounds were tested on the growth and antifeeding activity compared with toosendanin against fifth instar larvae Ostrinia furnacalis. The activities of two proteases, a weak alkaline trypsine like enzyme and a chymotrypsin like enzyme, in the midgut of Ostrinia furnacalis larvae were also measured. Experimental results suggest that when incorporated into an artificial diet at the concentration of 500mg/kg, the antifeeding activities of toosendanin,C 19 ,C 23 ,C 24 ,C 26 ,C 28 were 51 16%,57 61%,4 28%,51 08%,36 73% and 51 67%, respectively, C 19 ,C 24 ,C 28 had no significant difference with toosendanin. At 20mg/kg, the larval growth were remarkably suppressed by C 19 ,C 26 ,C 28 , the inhibition of C 28 was close to toosendanin in 48 h. The two proteases were activated by toosendanin and C 28 while they were inhibited in 48 h but activated in 24 h by C 19 ,C 24 and C 26 . In this paper, the related functions and mechanisms were discussed.展开更多
Excessive amounts of nitrate have accumulated in many soils on the North China Plain due to the large amounts of chemical N fertilizers or manures used in combination with low carbon inputs. We investigated the potent...Excessive amounts of nitrate have accumulated in many soils on the North China Plain due to the large amounts of chemical N fertilizers or manures used in combination with low carbon inputs. We investigated the potential of different carbon substrates added to transform soil nitrate into soil organic N （SON）. A 56-d laboratory incubation experiment using the 15N tracer （K15NO3） technique was carried out to elucidate the proportion of SON derived from accumulated soil nitrate following amendment with glucose or maize straw at controlled soil temperature and moisture. The dynamics and isotopic abundance of mineral N （NO3 and NH4＋） and SON and greenhouse gas （N20 and CO2） emissions during the incubation were investigated. Although carbon amendments markedly stimulated transformation of nitrate to newly formed SON, this was only a substitution effect of the newly formed SON with native SON because SON at the end of the incubation period was not significantly different （P 〉 0.05） from that in control soil without added C. At the end of the incubation period, amendment with glucose, a readily available C source, increased nitrate immobilization by 2.65 times and total N20-N emission by 33.7 times, as compared with maize straw amendment. Moreover, the differences in SON and total N20-N emission between the treatments with glucose and maize straw were significant （P 〈 0.05）. However, the total N20-N emission in the straw treatment was not significantly （P ~ 0.05） greater than that in the control. Straw amendment may be a potential option in agricultural practice for transformation of nitrate N to SON and minimization of N20 emitted as well as restriction of NO3-N leaching.展开更多
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.
文摘The genetic transformation was achieved by pollen_mediated approach on maize ( Zea mays L.) inbred lines Tai 9101 and Zong 31. Plasmid DNA of pGLⅡ_RC_1 was mixed with fresh pollen of maize inbreds in sucrose solution. The pollens were treated by ultrasonication and collected, pollinated on silks of maize ears. Transformants were confirmed by dot blot hybridization, PCR amplification and PCR_Southern blot hybridization. The pollen_mediated transformation approach could circumvent the tedious tissue culture procedures like in particle bombardment and Agrobacterium infection, etc. This approach is simple, easy to operate, and could be widely used in practice.
基金Supported by the Key Technology R&D Program of Tianjin(No.13ZCZDNC00900)the Tianjin Research Program of Application Foundation and Advanced Technology(No.14JCQNJC15100)+2 种基金the Tianjin Innovative Research Team(No.TD12-5018)the Tianjin Key Laboratory of Animal and Plant Resistancethe National Natural Science Foundation of China(No.31402313)
文摘A 9-week feeding trial in floating freshwater cages(1.0 m×1.0 m×2.0 m) was conducted to study the effects of different dietary levels of protein and starch on growth,body composition,and gene expression of enzymes in common carp,Cyprinus carpio(mean body weight,36.12±1.18 g) to evaluate the protein-sparing effect of dietary carbohydrate.Four diets were formulated with corn starch as the carbohydrate source to obtain corn starch levels of 6.5%,13%,19.5%,or 26%and protein levels of 30.5%,28.2%,26.4%,and 24.2%.The results showed no differences in growth performance of fish fed the diets with different protein and corn starch levels,but body composition and glucose metabolic enzyme activity of carp were significantly affected by the different diets(P<0.05).Weight gain,specific growth rate,and the feed conversion ratio were not different in fish fed the different dietary treatments.Protein efficiency ratio increased significantly as corn starch level increased(P<0.05).Whole-body crude lipid composition increased with increasing dietary corn starch level(P<0.05).Glucokinase(GK),hexokinase,and pyruvate kinase(PK) activities increased significantly with increasing dietary corn starch level(P<0.05),whereas glucose-6-phosphate(G6Pase) activity decreased with increasing dietary corn starch level(P<0.05).GK gene expression was significantly higher in fish fed the high-corn starch diet than those fed the low-corn starch diet(P<0.05).G6 pase gene expression tended to decrease with increasing starch level(P>0.05).In summary,the results indicate a protein-sparing effect by substituting carbohydrate in the diet of common carp.
文摘Five new compounds were tested on the growth and antifeeding activity compared with toosendanin against fifth instar larvae Ostrinia furnacalis. The activities of two proteases, a weak alkaline trypsine like enzyme and a chymotrypsin like enzyme, in the midgut of Ostrinia furnacalis larvae were also measured. Experimental results suggest that when incorporated into an artificial diet at the concentration of 500mg/kg, the antifeeding activities of toosendanin,C 19 ,C 23 ,C 24 ,C 26 ,C 28 were 51 16%,57 61%,4 28%,51 08%,36 73% and 51 67%, respectively, C 19 ,C 24 ,C 28 had no significant difference with toosendanin. At 20mg/kg, the larval growth were remarkably suppressed by C 19 ,C 26 ,C 28 , the inhibition of C 28 was close to toosendanin in 48 h. The two proteases were activated by toosendanin and C 28 while they were inhibited in 48 h but activated in 24 h by C 19 ,C 24 and C 26 . In this paper, the related functions and mechanisms were discussed.
基金Project supported by the National Natural Science Foundation of China(NSFC)(Nos.31172033 and 41101277)the National Science Basic Research Program of China(No.2007CB109308)+2 种基金the Foundation of the Chinese Ministry of Education for Ph.D.Work(No.20100008110004)the German Research Foundation (DFG)(No.IRTG 1070)the Innovation Group Grant of the National Natural Science Foundation of China(No.31121062)
文摘Excessive amounts of nitrate have accumulated in many soils on the North China Plain due to the large amounts of chemical N fertilizers or manures used in combination with low carbon inputs. We investigated the potential of different carbon substrates added to transform soil nitrate into soil organic N （SON）. A 56-d laboratory incubation experiment using the 15N tracer （K15NO3） technique was carried out to elucidate the proportion of SON derived from accumulated soil nitrate following amendment with glucose or maize straw at controlled soil temperature and moisture. The dynamics and isotopic abundance of mineral N （NO3 and NH4＋） and SON and greenhouse gas （N20 and CO2） emissions during the incubation were investigated. Although carbon amendments markedly stimulated transformation of nitrate to newly formed SON, this was only a substitution effect of the newly formed SON with native SON because SON at the end of the incubation period was not significantly different （P 〉 0.05） from that in control soil without added C. At the end of the incubation period, amendment with glucose, a readily available C source, increased nitrate immobilization by 2.65 times and total N20-N emission by 33.7 times, as compared with maize straw amendment. Moreover, the differences in SON and total N20-N emission between the treatments with glucose and maize straw were significant （P 〈 0.05）. However, the total N20-N emission in the straw treatment was not significantly （P ~ 0.05） greater than that in the control. Straw amendment may be a potential option in agricultural practice for transformation of nitrate N to SON and minimization of N20 emitted as well as restriction of NO3-N leaching.
