Along with the increase in the population,the requirements for increased human food and animal feed production have been boosted worldwide.Such an urgent food demand requires more arable land for crop productions.To c...Along with the increase in the population,the requirements for increased human food and animal feed production have been boosted worldwide.Such an urgent food demand requires more arable land for crop productions.To cope with these urgent needs,saline and/or alkaline lands could be used as alternatives for crop production.展开更多
Developing expressed sequence tag-derived SSR （EST-SSR） markers is imperative in genetic research. In this paper, we reported 37 EST-SSR markers which were developed from 286 unigenes obtained from soybean cDNA libr...Developing expressed sequence tag-derived SSR （EST-SSR） markers is imperative in genetic research. In this paper, we reported 37 EST-SSR markers which were developed from 286 unigenes obtained from soybean cDNA library. Among the 286 markers designed for the 4 accessions of Glycine max and 6 of its wild progenitor （G. soja） within the subgenus Soja, 209 markers amplified DNA fragments, taking 73.1% and 37 markers appeared to be polymorphic, which was 12.9% of the total. The 37 loci detected a total of 142 alleles, while the PIC values varied from 0.194 to 0.794. Both the number of alleles per locus and PIC value were significantly related to the SSR motif. Six EST-SSR loci may be fixed for different alleles between G. max and G. soja since they were particularly polymorphic among the 6 G. soja accessions. A neighbor-joining tree placed the G. max accessions together as a group within the G. soja, though the average genetic distance among G. soja accessions was much higher. These new EST-SSRs markers will be useful for genetic diversity analysis, genetic mapping construction and gene discovery in Soja subgenus.展开更多
Transgenic lines were achieved by transforming the E. coli 1-phosphate mannitol dehydrogenase gene （mtl-D） into the Populus tomentosa Cart. genome. An Agrobacterium tumefaciens strain （AGL1）, constructed by clonin...Transgenic lines were achieved by transforming the E. coli 1-phosphate mannitol dehydrogenase gene （mtl-D） into the Populus tomentosa Cart. genome. An Agrobacterium tumefaciens strain （AGL1）, constructed by cloning mtl-D into the disarmed plasmid pBin438, was used to infect leaves of the clone YW2. The infected leaf discs were cultured on a medium containing 30 mg.L 1 kanamycin and 500 mg.L 1 cefotaxime. Transgenic plantlets regenerated from the infected leaves, rooted on the medium con- taining 30 mg.L 1 kanamycin. PCR and a Southern blotting test verified that the exogenous mtl-D gene had integrated into the trans- formation plants of the P. tomentosa genome. The mannitol content in control plant was 69 gg.gl FW, and the mannitol contents of the transgenic lines T1 to T5 ranged between 103.7 and 289.5 μg·g^-1 FW. Of the shoots of the control plants 20% survived; on the medium containing 0.6% NaCl, 60% and 70% of two transgenic shoots survived on a medium containing 0.8% NaCI.展开更多
Different types of explants of China Rose （Rosa chinensis Jacq.） were placed on a Schenk and Hildebrandt （SH） medium containing L-proline and 2,4-dichlorophenoxyacetic acid （2,4-D）. Organogenesis was observed on...Different types of explants of China Rose （Rosa chinensis Jacq.） were placed on a Schenk and Hildebrandt （SH） medium containing L-proline and 2,4-dichlorophenoxyacetic acid （2,4-D）. Organogenesis was observed on callus induced from both whole leaf and petiole and the high frequency of organogenesis was observed on the whole leaf. Shoot regeneration was obtained via or- ganogenesis. The effects of pH and concentrations of antibiotics on maintenance of organogenesis capacity were investigated in sub- sequent subcultures. The pH value was found to play a critical role in retaining organogenesis capacity. The binary vector pBI121, carrying the gus gene coding for fl-glucuronidase （GUS） and the npt II gene mediated by Agrobacterium tumefaciens, was used for transformation of organogenic callus using 50 mg·L^-1 geneticin for selection. Six regenerated lines showed GUS activity, of which five were verified for the presence ofnpt Ⅱ gene by PCR.展开更多
The AhDREB1 gene, cloned from Atriplex hortensis L., was transferred into black locust （Robiniapseudoacacia L.） by an Agrobacterium-mediated transformation. The results suggest that stems of black locust sub-culture...The AhDREB1 gene, cloned from Atriplex hortensis L., was transferred into black locust （Robiniapseudoacacia L.） by an Agrobacterium-mediated transformation. The results suggest that stems of black locust sub-cultured in vitro for 20 d are suitable for genetic transformation. The optimum concentrations of kanamycin and cefotaxime were 30 and 150 mg.L-1, respectively. Important factors affecting the transformation efficiency were studied by means of a L9（3^4） orthogonal design. An effective system for genetic transformation in black locust was developed as follows： the stems were pre-cultured for 2 d, immersed in the Agrobacterium solution （OD600 = 0.7） with 10 mg·L^-1 acetosyringone for 21 min and then co-cultured for 2 d. The selection pressures, changing from low to high, could improve transformation efficiency. The transgenic plants were identified by a PCR method. The PCR results indicated that the AhDREB1 gene had been integrated into the genome of black locust and two lines of the transgenic plants were obtained.展开更多
Robinia pseudoacacia ‘Idaho＇ （Robinia x ambigua ‘Idahoensis＇, R. pseudoacacia x R. viscosa） modified by a mtLD gene went through five lines and had characteristics of drought tolerance. Three stages of their mic...Robinia pseudoacacia ‘Idaho＇ （Robinia x ambigua ‘Idahoensis＇, R. pseudoacacia x R. viscosa） modified by a mtLD gene went through five lines and had characteristics of drought tolerance. Three stages of their micropropagation had been studied by pre- vious investigators. The other two stages, in vitro shoot rooting and plantlet acclimatization, still remained unsolved in the laboratory. For this paper, we studied the later two stages based on the previous achievements. Results showed that the highest rooting rate of Idaho locust was 98.4% when the in vitro shoots, over 2.5 cm in height and 0.08 cm in diameter, were placed on a half strength MS basal medium with 0.4 mg.U1 IBA and 0.1 mg＇U1 NAA as supplements and were solidified with 0.5% agar; the highest survival rate was 98.3% when the rooted plantlets were potted in vermiculite. All the stages for micropropagation of the Idaho locust, modified by the mtl-D gene, were assembled completely. The tissue culture plants grow well in the field.展开更多
Robinia pseudoacacia ‘Idaho＇ is one of several multi-purpose trees used in ornamental, soil and water conservation, fodder and nectar sources. Plant abiotic stress tolerance transformed by genes could meet the requi...Robinia pseudoacacia ‘Idaho＇ is one of several multi-purpose trees used in ornamental, soil and water conservation, fodder and nectar sources. Plant abiotic stress tolerance transformed by genes could meet the requirements for reclamation of arid or alkalid lands and vegetation restoration. For this paper, we studied the effects of auxin and cytokine on Idaho locust in vitro regeneration and the establishment of gene transformation systems for plants mediated by Agrobacterium tumefaciens. Results showed that the ratios of cytokinin and auxin were the major factors affecting adventitious bud differentiation on a MS medium; the concentration of 0.5 mg·L^-16-BA benefitted callus proliferation and 0.25 mg·L^-1 IBA promoted shoot rooting; however, a higher IBA concentration will inhibit rooting. The most effective antitoxin for screening transgenic Idaho locust shoots was G418 and the most sensitive concentration of it was 8 mg·L^-1.展开更多
Based on the plant regeneration system, a GUS gene transformation system to Idaho locust （Robinia pseudoacacia ‘Idaho＇） mediated by Agrobacterium tumefaciens was established. The successful transformation was conf...Based on the plant regeneration system, a GUS gene transformation system to Idaho locust （Robinia pseudoacacia ‘Idaho＇） mediated by Agrobacterium tumefaciens was established. The successful transformation was confirmed by regenerating the shoots fi＇om the infected leaves in the presence of hygromysin; by histochemical X-gluc assays of 15-glucuronidase （GUS） and by PCR and PCR-Southern blotting analysis. The ratio of positive transgenic plants is 5.8% （5 out of 86 plants）. With this system, the target gene DREB was introduced into the leaves of Idaho locust. The transgenic plants regenerated, which was verified by PCR-Southern blot- ting. It is suggested that the transformation system could be a new, simple, reliable and practical route to gene transformation of R. pseudoacacia ＇Idaho＇ mediated with A. tumefaciens.展开更多
文摘Along with the increase in the population,the requirements for increased human food and animal feed production have been boosted worldwide.Such an urgent food demand requires more arable land for crop productions.To cope with these urgent needs,saline and/or alkaline lands could be used as alternatives for crop production.
基金supported by the National High-Tech R&D Program of China(863Program,2006AA10A110 and 2006AA10Z164)National Basic Research Program of China(973 Program,2010CB125900 and 2004CB117203)the Academy and Institute Foundation for Basic Scientific Research in Institute of Crop Science,Chinese Academy of Agricultural Sciences
文摘Developing expressed sequence tag-derived SSR （EST-SSR） markers is imperative in genetic research. In this paper, we reported 37 EST-SSR markers which were developed from 286 unigenes obtained from soybean cDNA library. Among the 286 markers designed for the 4 accessions of Glycine max and 6 of its wild progenitor （G. soja） within the subgenus Soja, 209 markers amplified DNA fragments, taking 73.1% and 37 markers appeared to be polymorphic, which was 12.9% of the total. The 37 loci detected a total of 142 alleles, while the PIC values varied from 0.194 to 0.794. Both the number of alleles per locus and PIC value were significantly related to the SSR motif. Six EST-SSR loci may be fixed for different alleles between G. max and G. soja since they were particularly polymorphic among the 6 G. soja accessions. A neighbor-joining tree placed the G. max accessions together as a group within the G. soja, though the average genetic distance among G. soja accessions was much higher. These new EST-SSRs markers will be useful for genetic diversity analysis, genetic mapping construction and gene discovery in Soja subgenus.
文摘Transgenic lines were achieved by transforming the E. coli 1-phosphate mannitol dehydrogenase gene （mtl-D） into the Populus tomentosa Cart. genome. An Agrobacterium tumefaciens strain （AGL1）, constructed by cloning mtl-D into the disarmed plasmid pBin438, was used to infect leaves of the clone YW2. The infected leaf discs were cultured on a medium containing 30 mg.L 1 kanamycin and 500 mg.L 1 cefotaxime. Transgenic plantlets regenerated from the infected leaves, rooted on the medium con- taining 30 mg.L 1 kanamycin. PCR and a Southern blotting test verified that the exogenous mtl-D gene had integrated into the trans- formation plants of the P. tomentosa genome. The mannitol content in control plant was 69 gg.gl FW, and the mannitol contents of the transgenic lines T1 to T5 ranged between 103.7 and 289.5 μg·g^-1 FW. Of the shoots of the control plants 20% survived; on the medium containing 0.6% NaCl, 60% and 70% of two transgenic shoots survived on a medium containing 0.8% NaCI.
文摘Different types of explants of China Rose （Rosa chinensis Jacq.） were placed on a Schenk and Hildebrandt （SH） medium containing L-proline and 2,4-dichlorophenoxyacetic acid （2,4-D）. Organogenesis was observed on callus induced from both whole leaf and petiole and the high frequency of organogenesis was observed on the whole leaf. Shoot regeneration was obtained via or- ganogenesis. The effects of pH and concentrations of antibiotics on maintenance of organogenesis capacity were investigated in sub- sequent subcultures. The pH value was found to play a critical role in retaining organogenesis capacity. The binary vector pBI121, carrying the gus gene coding for fl-glucuronidase （GUS） and the npt II gene mediated by Agrobacterium tumefaciens, was used for transformation of organogenic callus using 50 mg·L^-1 geneticin for selection. Six regenerated lines showed GUS activity, of which five were verified for the presence ofnpt Ⅱ gene by PCR.
基金supported by the Key Extended Project of State Forestry Administration, China (Grant No. 2003-5-2).
文摘The AhDREB1 gene, cloned from Atriplex hortensis L., was transferred into black locust （Robiniapseudoacacia L.） by an Agrobacterium-mediated transformation. The results suggest that stems of black locust sub-cultured in vitro for 20 d are suitable for genetic transformation. The optimum concentrations of kanamycin and cefotaxime were 30 and 150 mg.L-1, respectively. Important factors affecting the transformation efficiency were studied by means of a L9（3^4） orthogonal design. An effective system for genetic transformation in black locust was developed as follows： the stems were pre-cultured for 2 d, immersed in the Agrobacterium solution （OD600 = 0.7） with 10 mg·L^-1 acetosyringone for 21 min and then co-cultured for 2 d. The selection pressures, changing from low to high, could improve transformation efficiency. The transgenic plants were identified by a PCR method. The PCR results indicated that the AhDREB1 gene had been integrated into the genome of black locust and two lines of the transgenic plants were obtained.
文摘Robinia pseudoacacia ‘Idaho＇ （Robinia x ambigua ‘Idahoensis＇, R. pseudoacacia x R. viscosa） modified by a mtLD gene went through five lines and had characteristics of drought tolerance. Three stages of their micropropagation had been studied by pre- vious investigators. The other two stages, in vitro shoot rooting and plantlet acclimatization, still remained unsolved in the laboratory. For this paper, we studied the later two stages based on the previous achievements. Results showed that the highest rooting rate of Idaho locust was 98.4% when the in vitro shoots, over 2.5 cm in height and 0.08 cm in diameter, were placed on a half strength MS basal medium with 0.4 mg.U1 IBA and 0.1 mg＇U1 NAA as supplements and were solidified with 0.5% agar; the highest survival rate was 98.3% when the rooted plantlets were potted in vermiculite. All the stages for micropropagation of the Idaho locust, modified by the mtl-D gene, were assembled completely. The tissue culture plants grow well in the field.
文摘Robinia pseudoacacia ‘Idaho＇ is one of several multi-purpose trees used in ornamental, soil and water conservation, fodder and nectar sources. Plant abiotic stress tolerance transformed by genes could meet the requirements for reclamation of arid or alkalid lands and vegetation restoration. For this paper, we studied the effects of auxin and cytokine on Idaho locust in vitro regeneration and the establishment of gene transformation systems for plants mediated by Agrobacterium tumefaciens. Results showed that the ratios of cytokinin and auxin were the major factors affecting adventitious bud differentiation on a MS medium; the concentration of 0.5 mg·L^-16-BA benefitted callus proliferation and 0.25 mg·L^-1 IBA promoted shoot rooting; however, a higher IBA concentration will inhibit rooting. The most effective antitoxin for screening transgenic Idaho locust shoots was G418 and the most sensitive concentration of it was 8 mg·L^-1.
文摘Based on the plant regeneration system, a GUS gene transformation system to Idaho locust （Robinia pseudoacacia ‘Idaho＇） mediated by Agrobacterium tumefaciens was established. The successful transformation was confirmed by regenerating the shoots fi＇om the infected leaves in the presence of hygromysin; by histochemical X-gluc assays of 15-glucuronidase （GUS） and by PCR and PCR-Southern blotting analysis. The ratio of positive transgenic plants is 5.8% （5 out of 86 plants）. With this system, the target gene DREB was introduced into the leaves of Idaho locust. The transgenic plants regenerated, which was verified by PCR-Southern blot- ting. It is suggested that the transformation system could be a new, simple, reliable and practical route to gene transformation of R. pseudoacacia ＇Idaho＇ mediated with A. tumefaciens.
